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fg(:v*Q#  9$  qGACNNNNNNGTC,CCAGGCCTGG,GATCCGGATC,CCWGGTACCWGG,CCANNNNNTGG,CCAGGNNNTGG,CCANNNCCTGG,CCAGGNCCTGG,GCACNNNAACGTTNNNGTGC,CACNNNGTG,CCNNNNNNNGG,AACNAACRYGTGCGTGC,GCACGCACRYGTTNGTT,GCACNAACRYGTTNGTGC,AACGCACRYGTGCGTT,GACNNNNNGTC,GAANNNNNNNTTGG,CCAANNNNNNNTTC,CACNNNNGTG,AACACNNNNGTGC,GCACNNNNGTGTT,GCANNNNNNTGC,GAACNNNNNNTCC,GGANNNNNNGTTC,CAGNNNCTG,CCAGGNNCTG,CAGNNCCTGG,CCWGGGCCC,GGGCCCWGG,CCWGGGCCCWGG,AACNNNNNNGTGCACNNNNNNGTT,GACNNNGTC,GAANNNNTTC,CCWGGTACC,GGTACCWGG,CCWGGWCCWGG,ACNNNNGTAYC,GRTACNNNNGT,CCWGGYRCCWGG,GAAGNNNNNNTAC,GTANNNNNNCTTC,AACGGCNNNGTGC,GCACNNNGCCGTT,GCACGGCNNNGTT,AACNNNGCCGTGC,CGANNNNNNTGC,GCANNNNNNTCG,CGATCNNNNTGC,GCANNNNGATCG,TGANNNNNNTCA,TGATCNNNNTCA,TGANNNNGATCA,TGATCNNGATCA,AACCTGCNNGTGC,GCACNNGCAGGTT,GCACCTGCNNGTT,AACNNGCAGGTGC,GCCNNNNNGGC,AACNNNNNNGTGCMC,GKGCACNNNNNNGTT,CCWGGNCCWGG,AACTGGGNNGTGC,GCACNNCCCAGTT,GCACTGGGNNGTT,AACNNCCCAGTGC,GACNNNNGTC,GAGNNNNNCTC,CCWGGTCTC,GAGACCWGG,GATNNNNATC,GATCNNNATC,GATNNNGATC,GATCNNGATC,CCWGGCGCCWGG,ACNNNNNCTCC,GGAGNNNNNGT,AACNNNNNNGTGCAG,CTGCACNNNNNNGTT,GATCATGATC,CCWGGNNCCWGG,AACTGGNNNGTGC,GCACNNNCCAGTT,GCACTGGNNNGTT,AACNNNCCAGTGC,AACNAACCGGTGCGTGC,GCACGCACCGGTTNGTT,GCACNAACCGGTTNGTGC,AACGCACCGGTGCGTT,GCANNNNNTGC,GCACNNNNNNGTTCGAACNNNNNNGTGC,CCTNNNNNAGG,GCNNNNNNNGC,CCANNNNNNTGG,CCAGGNNCCTGG,CCTGGCCAGG,CAANNNNNGTGG,CCACNNNNNTTG,TTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAA,GCACNNNNNNGTTTAAACNNNNNNGTGC,CCAGGNCCY,RGGNCCTGG,CCWGGGNCCTGG,CCAGGNCCCWGG,AAGNNNNNCTT,GAYNNNNNVTC,GABNNNNNRTC,GAYNNNNNRTC,GATCNNNNVTC,GABNNNNGATC,GAYNNNNGATC,GATCNNNNRTC,GATCNNNGATC,GTYAACNNNNNNGTGC,GCACNNNNNNGTTRAC,GCACNNNNNNGTTAACNNNNNNGTGC,GATCNGATC,TAACTATAACGGTCCTAAGGTAGCGA,TCGCTACCTTAGGACCGTTATAGTTA,CTCTCTTAAGGTAGC,GCTACCTTAAGAGAG,TAGGGATAACAGGGTAAT,ATTACCCTGTTATCCCTA,GTTAACNNNNNNGTGC,GCACNNNNNNGTTAAC,AACNAACGCGTGCGTGC,GCACGCACGCGTTNGTT,GCACNAACGCGTTNGTGC,AACGCACGCGTGCGTT,TCCAACNNNNNNGTGC,GCACNNNNNNGTTGGA,CAYNNNNRTG,AACAYNNNNGTGC,GCACNNNNRTGTT,GTTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAAC,CCWGGCGCC,GGCGCCWGG,CCWGGNNCC,GGNNCCWGG,GATCGCGATC,AACNAACATGTGCGTGC,GCACGCACATGTTNGTT,GCACNAACATGTTNGTGC,AACGCACATGTGCGTT,TGGCAAACAGCTATTATGGGTATTATGGGT,ACCCATAATACCCATAATAGCTGTTTGCCA,ATCTATGTCGGGTGCGGAGAAAGAGGTAATGAAATGG,CCATTTCATTACCTCTTTCTCCGCACCCGACATAGAT,AACNNGCACGTGCNNGTT,GAACNNNNNCTC,GAGNNNNNGTTC,CCAGGWCCY,RGGWCCTGG,CCWGGGWCCTGG,CCAGGWCCCWGG,CCAGGWCCTGG,TTATAACNNNNNNGTGC,GCACNNNNNNGTTATAA,GCACNNNNNNGTTATAACNNNNNNGTGC,GAACNNNNNNTAC,GTANNNNNNGTTC,GGCCNNNNNGGCC,AACNNACTAGTGC,GCACTAGTNNGTT,AACTAGTNNGTGC,GCACNNACTAGTT,AACNAACTAGTGCGTGC,GCACGCACTAGTTNGTT,GCACNAACTAGTTNGTGC,AACGCACTAGTGCGTT,AACNNNAACGTGCNNGTGC,GCACNNGCACGTTNNNGTT,GCACCCANNNGTT,AACNNNTGGGTGC,TTAACNNNNNNGTGC,GCACNNNNNNGTTAA,CACNNNNNNTCC,GGANNNNNNGTG,CACNNNNGATCC,GGATCNNNNGTG,GATCTAGATC,CCANNNNNNNNNTGG,CCAGGCCT,AGGCCTGG,CCTCGAGG,GATCCGGA,TCCGGATC,GGCGCGCC,GCGATCGC,CCWGGWCC,GGWCCWGG,GATCGATC,GATCATGA,TCATGATC,GCGGCCGC,CCTGGCCR,YGGCCAGG,CCWGGNCC,GGNCCWGG,GGCCGGCC,RTGCGCAY,GATCNNGA,TCNNGATC,CGCGCGCG,CCTGGCCA,TGGCCAGG,CGCCGGCG,GTTTAAAC,GATCGCGA,TCGCGATC,TTAATTAA,VCTCGAGB,CCTGCAGG,CRCCGGYG,CGTCGACG,ATTTAAAT,GCCCGGGC,GATCTAGA,TCTAGATC,CACCTGC,GCAGGTG,CCTNAGG,GATCGAT,ATCGATC,CCTCAGC,GCTGAGG,GCTNAGC,CCTNAGC,GCTNAGG,GCTCTTC,GAAGAGC,GGTNACC,CGGWCCG,ACCWGGT,RGGNCCY,CCTGGAG,CTCCAGG,GGTGATC,GATCACC,GATCNGA,TCNGATC,GGGWCCC,GAAGATC,GATCTTC,CCCWGGG,TCCNGGA,TCCWGGA,RGGWCCY,TTATAA,AGGCCT,GACGTC,GTMKAC,TCCGGA,TGCGCA,ACCTGC,GCAGGT,GGTACC,AGTACT,GGYRCC,CCGCTC,GAGCGG,AACGTT,YGGCCR,RAATTY,CTGAAG,CTTCAG,CACGTG,GRCGYC,AGCGCT,CTTAAG,ACRYGT,ACCGGT,ACTAGT,CACGTC,GACGTG,GWGCWC,GTGCAC,CYCGRG,GGGCCC,ATTAAT,CCTAGG,TTCGAA,GCTAGC,GKGCMC,TGGCCA,GGATCC,GRGCYC,ATCGAT,CACGAG,CTCGTG,GGCGCC,GAAGAC,GTCTTC,GCATGC,GTATCC,GGATAC,TGATCA,ACTGGG,CCCAGT,CTRYAG,RGCGCY,AGATCT,GGNNCC,CTGGAG,CTCCAG,CTTGAG,CTCAAG,CGATCG,GGTCTC,GAGACC,YACGTR,CCNNGG,GAATGC,GCATTC,WCCGGW,GCAATG,CATTGC,RCCGGY,GCGCGC,GAGGAG,CTCCTC,CGGCCG,CCCAGC,GCTGGG,GTGCAG,CTGCAC,CGRYCG,CGTACG,CGTCTC,GAGACG,CCATGG,TCGCGA,GDGCHC,TGTACA,TCATGA,ACATGT,GTATAC,CCWWGG,CTCTTC,GAAGAG,GCNNGC,CCRYGG,GTTAAC,RCATGY,TACGTA,RGATCY,GCGATG,CATCGC,GCAGTG,CACTGC,CCCGGG,CCGCGG,TTTAAA,GGCGGA,TCCGCC,GAGCTC,GATATC,CTGRAG,CTYCAG,CAGCAG,CTGCTG,GAATTC,ATGCAT,CATATG,GTYRAC,AAGCTT,GTNNAC,TCNNGA,CAATTG,ACGCGT,TCCRAC,GTYGGA,GCCGGC,CMGCKG,GCCGAG,CTCGGC,CTCGAG,CTGCAG,CAGCTG,GTCGAC,CTYRAG,AATATT,GATCGA,TCGATC,GACCGA,CACCCA,TCGGTC,TGGGTG,WGTACW,TARCCA,TGGYTA,TCTAGA,GGATC,GATCC,CCWGG,GTCTC,GAGAC,GCWGC,GGNCC,CCSGG,GGTGA,TCACC,GGWCC,GCAGC,GCTGC,CCATC,GATGG,ACGGC,GCCGT,GCNGC,CCNGG,GCATC,GATGC,ACTGG,CCAGT,GGATG,CATCC,CTCAG,CTGAG,GGGAC,GTCCC,ACNGT,CTNAG,CAGTG,CACTG,GACGC,GCGTC,CCCGC,GCGGG,GANTC,CGWCG,TCNGA,CCTTC,GAAGG,GTNAC,GAAGA,TCTTC,GAGTC,GACTC,GTSAC,GAWTC,GCSGC,CASTG,ATGAA,TTCAT,ACGGA,TCCGT,CGCG,CCGC,GCGG,GTAC,AGCT,GCGC,CTAG,GATC,GGCC,CCGG,CATG,RGCY,YATR,ACGT,TGCA,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                        .                                                                                                                                                                               	                                                                                                                                G                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     	                                                                                                      	                                  	                                                                              4                           .                                             	                                           8          
                                 	                   	                	      &                                                                                    %   	      *   )      
      
                     Q   5                                                	                                  
            '                  -   @   6      C   :      ,   C   $   )         !   5   S   *   .   V                        .                                                                                 	                                                            G                                                                                                                                                                                                                                                     	                                    	              	                            4         .               	               8    
           	       	      	  &                              % 	  * )  
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       Q 5                	            
    '      - @ 6  C :  , C $ )   ! 5 S * . V    Y  !  K    w    O!	i]3$` a    ]<=wXcw!	r	C

D6imxE1H PY !\#,	P
!Uwp!!J%<)*!$}2A'$!
a%'N*SxR;.!8rq*aU"\2r!;) Y p S'JS\xE.R[2;	1
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!LLl	
V\`+:eaelF p.I
Ul,pY,)Z!@%Z!.+" p46	k	X#c#,Y !,	[*.	X#1O A-
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X#"Z$/)Fm'?x')h,n(Y	\  [ O!L		L+,QZi~"]KxB>+t0r	lG M{$%C'(A(()*b+,,n
	$
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R & 'O?L,L	o9&~	?	OhRuk$?	O    ZR#,,#(G)p&] "?		E
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:rc6?pu)   !!S#!&5*Y  nuGX$vbhW6
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:
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u 0n
G !  Qr IY #$(())++8,; j
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R !&	 I!L!,[4y}!(+$xB V"&Q),  ?			xO!Xi#')h,B  n*=GU
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5U@Z,}%C&x	Z,,,v$&)T+j2m  (*d KeA##CPp`gb+	MW'!* l(4 ?&,X#B.Lp5}	I

:/cu)tY !!!1"T(+>IV+jY\$,- 0]7m>

{U+pf|cu !H!K!!%E&;)6 UmR1qZ+7	l	L
03Wf,1 oY#d#g*,,   T}*I	J	
K.	2#%'*6 q+0f1Y#g*- 7m
U+pfcu !H!K!!;)0]>
{|%E&	
/&f(\)v** rN@u5"		
g

0~C130- .
0].GvBE27R m>
y

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߸z,j 0O\0¾8>خɧϨHN<\|LS?Y_>2ƶ4t*x04sZX7zòbhZtӘ}35TNS.{K]\a)5uET#cƂ%'#5vͣDfH;\nYSHRH!	yKHVM LT}N4ã37D1 IUj9DI0U)CQUfVvdM9)o]PGVxNutEme$"6DeQPSFDMWU%|50GAfhHZfVeI%VI75DQ TM A:4Sy2t=i#M4]j8@3]6{tmM;[F~T];
;W}JVѨ`HDVZwS率EUUQiZFVRv]Z i$$*Qd'!dJC@flG@>jj6g8DeZZ]VrRYj	*4U*`ycz&j:Aw(gv9Vim}V몖^Yib5zXMVUW9*^̢Y9}诔Ӿ:ڪnb{N:6ui*]Պ7o]Z:NjNWZeu؆㪩ӫbykeXYgٵ7۩hEzŎ#c~D~uڊkj#ZuZ]Vխhmhjӫ$ۢefjfkjZYj˞LF~eYZ*BzM[aeZ/f$iuR֡)@;
kRHTf7enY.yb*mM>YNnNZvoee5WUo)5[ҕ*YfLkVފu^X2ZmiVjuX}*=Vnen6ةg٭L&mZj]V6;~\o%VVZ[룺'.Y5)VP伦סU :SyX4jEP h 'HEGe]^TZuWVUVDoz<;   !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  ^<?xml version="1.0"?><Features nextValidID="20"><Feature recentID="19" name="EF1a" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7-1265" color="#ffffff" type="standard"/><Q name="note"><V text="EF1a"/></Q></Feature><Feature recentID="18" name="5'UTR exon1" type="5'UTR" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="285-317" color="#a6acb3" type="standard"/><Q name="note"><V text="5'UTR exon1"/></Q></Feature><Feature recentID="17" name="Intron1" type="intron" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="318-1256" color="noColor" type="standard"/><Q name="note"><V text="Intron1"/></Q></Feature><Feature recentID="16" name="Puro" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1278-1877" color="#993366" type="standard"/><Q name="note"><V text="Puro"/></Q></Feature><Feature recentID="15" name="WPRE" type="misc_binding" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1893-2481" color="#a6acb3" type="standard"/><Q name="note"><V text="WPRE"/></Q></Feature><Feature recentID="14" name="3'LTR" type="3'UTR" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2552-2787" color="#a6acb3" type="standard"/><Q name="note"><V text="3'LTR"/></Q></Feature><Feature recentID="13" name="SV40 polyadenylation signal" type="polyA_signal" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2864-2994" color="#a6acb3" type="standard"/><Q name="note"><V text="SV40 polyadenylation signal"/></Q></Feature><Feature recentID="12" name="SV40 origin of replication" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3074-3149" color="#ffff00" type="standard"/><Q name="note"><V text="SV40 origin of replication"/></Q></Feature><Feature recentID="11" name="T7" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3180-3199" color="#ffffff" type="standard"/><Q name="note"><V text="T7"/></Q></Feature><Feature recentID="10" name="F1 ori" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3367-3822" color="#ffff00" type="standard"/><Q name="note"><V text="F1 ori"/></Q></Feature><Feature recentID="9" name="AmpR" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3953-4813" color="#993366" type="standard"/><Q name="note"><V text="AmpR"/></Q></Feature><Feature recentID="8" name="pUC Ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5074-5689" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="pUC Ori"/></Q></Feature><Feature recentID="7" name="5'LTR" type="misc_structure" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6037-6449" color="#a6acb3" type="standard"/><Q name="note"><V text="5'LTR"/></Q></Feature><Feature recentID="6" name="Psi" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6500-6637" color="#a6acb3" type="standard"/><Q name="note"><V text="Psi"/></Q></Feature><Feature recentID="5" name="gag" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6604-6968" color="#a6acb3" type="standard"/><Q name="note"><V text="gag"/></Q></Feature><Feature recentID="4" name="RRE" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7114-7355" color="#a6acb3" type="standard"/><Q name="note"><V text="RRE"/></Q></Feature><Feature recentID="3" name="hU6" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7753-8002" color="#ffffff" type="standard"/><Q name="note"><V text="hU6"/></Q></Feature><Feature recentID="2" name="Filler" type="misc_binding" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7998-9882" color="#a6acb3" type="standard"/><Q name="note"><V text="Filler"/></Q></Feature><Feature recentID="1" name="chRNA" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9883-9964" color="#a6acb3" type="standard"/><Q name="note"><V text="chRNA"/></Q></Feature><Feature recentID="0" name="CPPT" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="10014-10131" color="#a6acb3" type="standard"/><Q name="note"><V text="CPPT"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="reverse primer for deep seq in nature paper" sequence="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="10051-10095" boundStrand="1" annealedBases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC" meltingTemperature="62"><Component hybridizedRange="10051-10095" bases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC"/></BindingSite><BindingSite simplified="1" location="10051-10095" boundStrand="1" annealedBases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC" meltingTemperature="62"><Component hybridizedRange="10051-10095" bases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC"/></BindingSite></Primer><Primer recentID="1" name="forward primer for deep seq in nature paper" sequence="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7919-7959" boundStrand="0" annealedBases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG" meltingTemperature="62"><Component hybridizedRange="7919-7959" bases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG"/></BindingSite><BindingSite simplified="1" location="7919-7959" boundStrand="0" annealedBases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG" meltingTemperature="62"><Component hybridizedRange="7919-7959" bases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG"/></BindingSite></Primer></Primers>
  <Notes>
<UUID>5608483e-eca8-496b-b390-f067e4f2cadc</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Description>-</Description>
<Created UTC="5:0:0">2014.7.18</Created>
<LastModified UTC="23:0:22">2018.12.26</LastModified>
<AccessionNumber>-</AccessionNumber>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
<Comments>AKA pHKO_09 ApEinfo:methylated:1</Comments>
</Notes>
     	  _  4         4>30Z)O:<"2b}<!W.MAcPbPSJNuuˉ1CL߃=T"eߍPuH(dD`S&?E	h#v:,BW.C r
Uwg!5N+|J"@=SR}?!~p:݅4zRP/A3}Pq^uR_6@5Iםt\`EmBFv4ӳ}4]p#?}7TL݂J|m`q8Is)K  <AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>Unmodified</UpstreamModification><DownstreamModification>Unmodified</DownstreamModification></AdditionalSequenceProperties>
  	<?xml version="1.0"?><Features nextValidID="5"><Feature recentID="4" name="eCFP" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="598-1314" color="#00ccff" type="standard"/></Feature><Feature recentID="3" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="1315-1321" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="2" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="456-477" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="1" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="72-91" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="0" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="36-597" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Fragment.FOR (2)" sequence="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"><BindingSite location="0-34" boundStrand="0" annealedBases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG" meltingTemperature="74"><Component hybridizedRange="0-34" bases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"/></BindingSite><BindingSite simplified="1" location="0-34" boundStrand="0" annealedBases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG" meltingTemperature="74"><Component hybridizedRange="0-34" bases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"/></BindingSite></Primer><Primer recentID="1" name="Fragment.REV (2)" sequence="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"><BindingSite location="1295-1331" boundStrand="1" annealedBases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG" meltingTemperature="68"><Component hybridizedRange="1295-1331" bases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"/></BindingSite><BindingSite simplified="1" location="1295-1331" boundStrand="1" annealedBases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG" meltingTemperature="68"><Component hybridizedRange="1295-1331" bases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"/></BindingSite></Primer></Primers>
  g<Notes>
<UUID>96873316-6a85-4c84-994d-52ea033d7808</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:2:28">2018.12.26</Created>
<LastModified UTC="23:2:28">2018.12.26</LastModified>
<CreatedBy>Li Lab copy3</CreatedBy>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  A\     {  -         -hY)V)Zm;j}d<VUVE+Ҡ]D {-P1U1*#Sy[TƤN T1ELT985"L-3%:X93/kC׊@0>9=81PPƔ%@-(q3%{y1h	dZ~'R8UYOʏj㚥nڋZa*]eI'wid&!"Xzz]V(&l(=e5c,K_IN/tSn-\(MPۂҦYנ9TU)EiRv)YQY}Mj믊A-J





		-VQYjFW  ]ޔj>rTU%E[Y9n.i>nx㮫=?Et6#PM7X6./}ЗWrvtTn(u*!bzLTmhF֊D(4MQuU!
idWu]jnnvڹ689E/nr}c}h=9`kA`G/]Cq'>~(ж}Trn+GfwB
JtB%TZ!Ru'ZRU&[%RUR{UziAITMEE^Dk:U]aX*iytucw9RW$SU-OMo#S<cD"i.DCY4MwCрZQ91kUS|{(ej;tG'UQ5UZj)`]jY!mehBcja]U%oVZUg΅&cFiXF8tjYWaڢQ`V}jZm[sdU;J]HjUf%WxQ*TbZ{NDeFNY4CWUvGFE`#fޥz_EEiU}m]hzT(Ql`Xf=QZQ+?R~WZB(!n+UTIVTZUVfAVufdmݦu8V֘օjtZ١u9TZiT7V$]XBB67]9MR֕#]&UUv8U4mآZ*iuQEmNVN(SZU]U#]QWc)SNvF;UXhSA8|ԂiPuJyW*jکRUAvUkJiUjdYjb&˼"uSVgR%6aWeUWVzrj囨-ZZ&cVaAmVChy)[Zpi+d+̧eVσYaɊ\hr%x4"m'?Xpczz'nri-}O+&4 0   C 0@000*C0ULS3?;??øx|{B%΂ʺ !SP;@0 22""2 L#ÈL02#1 L#0 8uC"2 @087(7ì;2;< 0Ѐ0ϰ< 08=%13@znnL03AAX#,<O<Ï  0
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Uwg!5N+|J"@=SR}?!~p:݅4zRP/A3}Pq^uR_6@5Iםt\`EmBFv4ӳ}4]p#?}7TL݂J|m`q8IS1D.FmVaOn9eAZY"(Pv`5mWmj
*vcَ=9f/kdU>
Fj]jѪ{N/o~4NQ=o8j8ӎ[vzڮjZP#hYazAظ}jٖ!WeeڕxkZ
Y5VE!VRzxhUIꪹUXIThԕ%VUe%%Z{Rz~4oy+00;Ӫ+)h2[`N*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	yKw=Ox%i4LBbCD wAj	=)UNo]_|**54ѧ3EUAQF. \UPPnoVXR[[&fWt:[HC~nJ>,yp:}?8jm5P<LH/zϽ.ihS%$5(t6"철:Ӽ+1)o#US
N-H)MeQx!'-d1(-g|vMThr1WPn8ұ{#^6ꞭʸMUPcU@Y@pV(Yfڛ@vj&UeudڿR   !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (<?xml version="1.0"?><Features nextValidID="25"><Feature recentID="24" name="eCFP" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7666-8382" color="#00ccff" type="standard"/></Feature><Feature recentID="23" name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6958-6987" color="#a020f0" type="standard"/></Feature><Feature recentID="22" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="8383-8393" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="21" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7524-7545" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="20" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="7140-7159" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="19" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7104-7665" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature recentID="18" name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="10697-11488" color="#ccffcc" type="standard" translated="1"/><Segment name="signal sequence" range="11489-11557" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9938-10526" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5335-5714" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3081-3082" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5717-5992" color="#ffffff" type="standard"/></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="11558-11662" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8488-8543" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment name="-10" range="9584-9590" color="#ffffff" type="standard"/><Segment range="9591-9608" color="#ffffff" type="standard"/><Segment name="-35" range="9609-9614" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9629-9650" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9497-9515" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9444-9460" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9536-9552" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9560-9576" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="gRNA scaffold" directionality="2" type="misc_RNA" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="3127-3202" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="U6 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5092-5332" color="#ffffff" type="standard"/><Q name="label"><V text="U6 Promoter"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="stuffer" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3199-5087" color="#a6acb3" type="standard"/></Feature></Features>
  
m<?xml version="1.0"?><Primers nextValidID="4"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="Fragment.REV (2)" sequence="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"><BindingSite location="8363-8388" boundStrand="1" annealedBases="CGCTTACTTGTACAGCTCGTCCATG" meltingTemperature="59"><Component bases="TGTCGACTTAA"/><Component hybridizedRange="8386-8388" bases="CGC"/><Component bases="G"/><Component hybridizedRange="8363-8384" bases="TTACTTGTACAGCTCGTCCATG"/></BindingSite><BindingSite simplified="1" location="8363-8388" boundStrand="1" annealedBases="CGCTTACTTGTACAGCTCGTCCATG" meltingTemperature="59"><Component bases="TGTCGACTTAA"/><Component hybridizedRange="8386-8388" bases="CGC"/><Component bases="G"/><Component hybridizedRange="8363-8384" bases="TTACTTGTACAGCTCGTCCATG"/></BindingSite></Primer><Primer recentID="2" name="Fragment.FOR (2)" sequence="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"><BindingSite location="7083-7102" boundStrand="0" annealedBases="CTCGAGCGGGATCAATTCCG" meltingTemperature="60"><Component bases="CGGTGCCTGAACGCG"/><Component hybridizedRange="7083-7102" bases="CTCGAGCGGGATCAATTCCG"/></BindingSite><BindingSite simplified="1" location="7083-7102" boundStrand="0" annealedBases="CTCGAGCGGGATCAATTCCG" meltingTemperature="60"><Component bases="CGGTGCCTGAACGCG"/><Component hybridizedRange="7083-7102" bases="CTCGAGCGGGATCAATTCCG"/></BindingSite></Primer><Primer recentID="0" name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"><BindingSite location="3067-3109" boundStrand="0" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="73"><Component hybridizedRange="3067-3109" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite><BindingSite simplified="1" location="3067-3109" boundStrand="0" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="73"><Component hybridizedRange="3067-3109" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite></Primer><Primer recentID="1" name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"><BindingSite location="5302-5346" boundStrand="1" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="66"><Component hybridizedRange="5302-5346" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite><BindingSite simplified="1" location="5302-5346" boundStrand="1" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="66"><Component hybridizedRange="5302-5346" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite></Primer></Primers>
  k<Notes>
<UUID>4ffd57eb-e621-4411-9cbf-4ff8b35d0efb</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="5:0:0">2015.10.20</Created>
<LastModified UTC="23:2:28">2018.12.26</LastModified>
<CreatedBy>Tumor Cell Biology</CreatedBy>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
  9     
  *         *hY)V)Zm;j}d<VUVE+Ҡ]D {-P1U1*#Sy[TƤN T1ELT985"L-3%:X93/kC׊@0>9=81PPƔ%@-(q3%{y1h	dZ~'R8UYOʏj㚥nڋZa*]eI'wid&!"Xzz]V(&l(=e5c,K_IN/tSn-\(MPۂҦYנ9TU)EiRv)YQY}Mj믊A-J





		-VQYjFW  ]ޔj>rTU%E[Y9n.i>nx㮫=?Et6#PM7X6./}ЗWrvtTn(u*!bzLTmhF֊D(4MQuU!
idWu]jnnvڹ689E/nr}c}h=9`kA`G/]Cq'>~(ж}Trn+GfwB
JtB%TZ!Ru'ZRU&[%RUR{UziAITMEE^Dk:U]aX*iytucw9RW$SU-OMo#S<cD"i.DCY4MwCрZQ91kUS|{(ej;tG'UQ5UZj)`]jY!mehBcja]U%oVZUg΅&cFiXF8tjYWaڢQ`V}jZm[sdU;J]HjUf%WxQ*TbZ{NDeFNY4CWUvGFE`#fޥz_EEiU}m]hzT(Ql`Xf=QZQ+?R~WZB(!n+UTIVTZUVfAVufdmݦu8V֘օjtZ١u9TZiT7V$]XBB67]9MR֕#]&UUv8U4mآZ*iuQEmNVN(SZU]U#]QWc)SNvF;UXhSA8|ԂiPuJyW*jکRUAvUkJiUjdYjb&˼"uSVgR%6aWeUWVzrj囨-ZZ&cVaAmVChy)[Zpi+d+̧eVσYaɊ\hr%x4"m'?Xpczz'nri-}O+&4 0   C 0@000*C0ULS3?;??øx|{B%΂ʺ !SP;@0 22""2 L#ÈL02#1 L#0 8uC"2 @087(7ì;2;< 0Ѐ0ϰ< 08=%13@znnL03AAX#,<O<Ï  0
î|0ND_b>ꯝ;5࢈5E$0Z)O:<"2b}<!W.MAcPbPSJNuuˉ1CL߃=T"eߍPuH(dD`S&?E	h#v:,BW.C r
UwgL<ǹĹnZ`m?MzjjdZ"Aـ@ն]XI*D5ُf88T TBIWO8T)j)vUv*OFy88n1Fp; N8amjej-B8	db-bfX\jCEjV65h*	dYUYIVUPIU'UbZRU%Rhj{RT[UzRtԕiIw
SѽD|NH@3m8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x{8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%ˡ/_QzlM=A1	#9EI&;TJ	;5u}L FUE sqW)@jFUAߑZUaJmnTI^ҟXm!Ln.j({$e@HXgG;|;ѩW@3E 5o>NDҖJ(9 sʂ`kNPħa(UL);(RR"{`7cF:!ԶE&0ģ?Y=4^R;mU ^A|oJ%xn۪z*!5WACU5UeXg{hSjm.L٨XUՐ
7jH  '<AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>FivePrimePhosphorylated</UpstreamModification>
<DownstreamModification>FivePrimePhosphorylated</DownstreamModification>
</AdditionalSequenceProperties>

  'r<?xml version="1.0"?><Features recycledIDs="21,22,23,24,25,26" nextValidID="29"><Feature recentID="28" name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6958-6987" color="#a020f0" type="standard"/></Feature><Feature recentID="27" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="7666-7676" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="20" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7524-7545" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="19" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="7140-7159" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="18" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7104-7665" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature recentID="17" name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="9980-10771" color="#ccffcc" type="standard" translated="1"/><Segment name="signal sequence" range="10772-10840" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="16" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9221-9809" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5335-5714" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3081-3082" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5717-5992" color="#ffffff" type="standard"/></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="10841-10945" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7771-7826" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment name="-10" range="8867-8873" color="#ffffff" type="standard"/><Segment range="8874-8891" color="#ffffff" type="standard"/><Segment name="-35" range="8892-8897" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8912-8933" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8780-8798" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8727-8743" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8819-8835" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8843-8859" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="gRNA scaffold" directionality="2" type="misc_RNA" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="3127-3202" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="U6 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5092-5332" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="label"><V text="U6 Promoter"/></Q></Feature></Features>
  Q<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"><BindingSite location="3067-3109" boundStrand="0" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="72"><Component hybridizedRange="3067-3109" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite><BindingSite simplified="1" location="3067-3109" boundStrand="0" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="72"><Component hybridizedRange="3067-3109" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite></Primer><Primer recentID="1" name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"><BindingSite location="5302-5346" boundStrand="1" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="65"><Component hybridizedRange="5302-5346" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite><BindingSite simplified="1" location="5302-5346" boundStrand="1" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="65"><Component hybridizedRange="5302-5346" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite></Primer></Primers>
   <Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created>2015.10.20</Created>
<LastModified>2015.11.30</LastModified>
<CreatedBy>Tumor Cell Biology</CreatedBy>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
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*vcَ=9f/kdU>
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Y5VE!VRzxhUIꪹUXIThԕ%VUe%%Z{Rz~4oy+00;Ӫ+)h2[`N*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	yKw=Ox%i4LBbCD wAj	=)UNo]_|**54ѧ3EUAQF. \UPPnoVXR[[&fWt:[HC~nJ>,yp:}?8jm5P<LH/zϽ.ihS%$5(t6"철:Ӽ+1)o#US
N-H)MeQx!'-d1(-g|vMThr1WPn8ұ{#^6ꞭʸMUPcU@Y@pV(Yfڛ@vj&UeudڿR   '<AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>FivePrimePhosphorylated</UpstreamModification>
<DownstreamModification>FivePrimePhosphorylated</DownstreamModification>
</AdditionalSequenceProperties>

  3<?xml version="1.0"?><Features nextValidID="29"><Feature recentID="28" name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6958-6987" color="#a020f0" type="standard"/></Feature><Feature recentID="27" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="8361-8393" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="26" name="SJVL-498_Max=860" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="8332-8350" color="#00ff00" type="standard"/><Q name="note"><V text="GFP PCR right.  Max recorded sequencing read = 860  bases.  Perfect match Tm = 63.2 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  CGGCGGCGGT CACGAACTC.  This primer matches this site  completely."/></Q></Feature><Feature recentID="25" name="SJVL-403_Max=772" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="8210-8238" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing Primers for HRST.  Max recorded  sequencing read = 772 bases.  Perfect match Tm = 71.8 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GTCGCCGATG GGGGTGTTCT GCTGGTAGT.  This  primer matches this site completely."/></Q></Feature><Feature recentID="24" name="AAV5_TR_C" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8235-8242" color="#000000" type="standard"/><Q name="note"><V text="AAV serotype 5 ITR region C."/></Q></Feature><Feature recentID="23" name="SJVL-497_Max=794" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8065-8088" color="#00ff00" type="standard"/><Q name="note"><V text="GFP PCR left.  Max recorded sequencing read = 794  bases.  Perfect match Tm = 67 C, although for oligos less  than 35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  GACGGCAACA TCCTGGGGCA CAAG.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="22" name="SJVL-404_Max=836" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="7745-7770" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing Primers for HRST.  Max recorded  sequencing read = 836 bases.  Perfect match Tm = 73.4 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is CTCGCCCTCG CCGGACACGC TGAACT.  This  primer matches this site completely."/></Q></Feature><Feature recentID="21" name="mCherry" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7666-8382" color="#ff0000" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="EGFP from Clontech pEGFP-N1 and C1, such that both  polylinkers are excluded."/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="20" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7524-7545" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="19" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="7140-7159" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="18" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7104-7665" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature recentID="17" name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="10697-11488" color="#ccffcc" type="standard" translated="1"/><Segment name="signal sequence" range="11489-11557" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="16" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9938-10526" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5335-5714" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3081-3082" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5717-5992" color="#ffffff" type="standard"/></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="11558-11662" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8488-8543" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment name="-10" range="9584-9590" color="#ffffff" type="standard"/><Segment range="9591-9608" color="#ffffff" type="standard"/><Segment name="-35" range="9609-9614" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9629-9650" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9497-9515" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9444-9460" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9536-9552" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9560-9576" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="gRNA scaffold" directionality="2" type="misc_RNA" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="3127-3202" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="U6 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5092-5332" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="label"><V text="U6 Promoter"/></Q></Feature></Features>
  Q<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"><BindingSite location="3067-3109" boundStrand="0" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="72"><Component hybridizedRange="3067-3109" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite><BindingSite simplified="1" location="3067-3109" boundStrand="0" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="72"><Component hybridizedRange="3067-3109" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite></Primer><Primer recentID="1" name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"><BindingSite location="5302-5346" boundStrand="1" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="65"><Component hybridizedRange="5302-5346" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite><BindingSite simplified="1" location="5302-5346" boundStrand="1" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="65"><Component hybridizedRange="5302-5346" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite></Primer></Primers>
   <Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created>2015.10.20</Created>
<LastModified>2015.10.20</LastModified>
<CreatedBy>Tumor Cell Biology</CreatedBy>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
  ;+     	G  $         $hY)V)Zm;j}d<VUVE+Ҡ]D {-P1U1*#Sy[TƤN T1ELT985"L-3%:X93/kC׊@0>9=81PPƔ%@-(q3%{y1h	dZ~'R8UYOʏj㚥nڋZa*]eI'wid&!"Xzz]V(&l(=e5c,K_IN/tSn-\(MPۂҦYנ9TU)EiRv)YQY}Mj믊A-J





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idWu]jnnvڹ689E/nr}c}h=9`kA`G/]Cq'>~(ж}Trn+GfwB
JtB%TZ!Ru'ZRU&[%RUR{UziAITMEE^Dk:U]aX*iytucw9RW$SU-OMo#S<cD"i.DCY4MwCрY% O٘v^%`8|q%Y-` yWރIR&ُRs	X?>?I~v&9r{ۿZjjji#L`   0  1=mR`00eT30K8>>;̧t!00?ZL*Xp쫪2># 0 ##""# 20?803:"3< ?# @T8"#  !4 p: 0  3 <; LË1<>[#4V<V<S<4>23/n  8 p :ȇO+c뮪Ӿ\
(.jTZrH{?rYC#*.f)٣˪b?~ T5,4GW\441ЏOr&Q7_rJJ TKV2k]@8PcPn>7c؊)Ur?_p Wq}P1 81\=ԭ2/@cߜ58+n7N?#?#<}SMǮ/=r7ۮ5Q+1lNPA] NyOu˙M\6!zo=lAL;<}E8~K5Gqm/<6ԕ0KfѺUi6냖T5"j{fVڕva&f=]PQm&a]?ESzj*Vتy=Kd`FO#8ᅷg)е6%vTX]֭bqV]ZWLֵ%PdXRUe'YVA%TUiITIIRUoVYI[RU'߸+MOF=:!΃%d⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ.	Gq4]D&$0tJR>^py&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%zJ}c71+odȑgۃ#a1FY_OOש斊<9RJJY*3\hCl-*	;¿\mCآ5U0/lIJ4m킘46Z-o%긇{_RH2pgdzHW+{+7nL]V<TwbiM
ha0gfbjEVWV@+ݫU  '<AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>FivePrimePhosphorylated</UpstreamModification>
<DownstreamModification>FivePrimePhosphorylated</DownstreamModification>
</AdditionalSequenceProperties>

  .<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4704-4733" color="#a020f0" type="standard"/></Feature><Feature recentID="24" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="6107-6139" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="23" name="SJVL-498_Max=860" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="6078-6096" color="#00ff00" type="standard"/><Q name="note"><V text="GFP PCR right.  Max recorded sequencing read = 860  bases.  Perfect match Tm = 63.2 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  CGGCGGCGGT CACGAACTC.  This primer matches this site  completely."/></Q></Feature><Feature recentID="22" name="SJVL-403_Max=772" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="5956-5984" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing Primers for HRST.  Max recorded  sequencing read = 772 bases.  Perfect match Tm = 71.8 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GTCGCCGATG GGGGTGTTCT GCTGGTAGT.  This  primer matches this site completely."/></Q></Feature><Feature recentID="21" name="AAV5_TR_C" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="5981-5988" color="#000000" type="standard"/><Q name="note"><V text="AAV serotype 5 ITR region C."/></Q></Feature><Feature recentID="20" name="SJVL-497_Max=794" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="5811-5834" color="#00ff00" type="standard"/><Q name="note"><V text="GFP PCR left.  Max recorded sequencing read = 794  bases.  Perfect match Tm = 67 C, although for oligos less  than 35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  GACGGCAACA TCCTGGGGCA CAAG.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="19" name="SJVL-404_Max=836" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="5491-5516" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing Primers for HRST.  Max recorded  sequencing read = 836 bases.  Perfect match Tm = 73.4 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is CTCGCCCTCG CCGGACACGC TGAACT.  This  primer matches this site completely."/></Q></Feature><Feature recentID="18" name="mCherry" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5412-6128" color="#ff0000" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="EGFP from Clontech pEGFP-N1 and C1, such that both  polylinkers are excluded."/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="17" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="5270-5291" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="16" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="4886-4905" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="15" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4850-5411" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature recentID="14" name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="8443-9234" color="#ccffcc" type="standard" translated="1"/><Segment name="signal sequence" range="9235-9303" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7684-8272" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3081-3460" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3463-3738" color="#ffffff" type="standard"/></Feature><Feature recentID="9" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="9304-9408" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6234-6289" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment name="-10" range="7330-7336" color="#ffffff" type="standard"/><Segment range="7337-7354" color="#ffffff" type="standard"/><Segment name="-35" range="7355-7360" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7375-7396" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7243-7261" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7190-7206" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7282-7298" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7306-7322" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
   <?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  U<Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pT2K-CAGGS-IRES-mCherry</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Created>2014.3.27</Created>
<LastModified>2014.12.30</LastModified>
<CreatedBy>Trial User</CreatedBy>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
  $                hY)V)Zm;j}d<VUVE+Ҡ]D {-P1U1*#Sy[TƤN T1ELT985"L-3%:X93/kC׊@0>9=81PPƔ%@-(q3%{y1h	dZ~'R8UYOʏj㚥nڋZa*]eI'wid&!"Xzz]V(&l(=e5c,K_IN/tSn-\(MPۂҦYנ9TU)EiRv)YQY}Mj믊A-J





		-VQYjFW  ]ޔj>rTU%E[Y9n.i>nx㮫=?Et6#PM7X6./}ЗWrvtTn(u*!bzLTmhF֊D(4MQuU!
idWu]jnnvڹ689E/nr}c}h=9`kA`G/]Cq'>~(ж}Trn+GfwB
JtB%TZ!Ru'ZRU&[%RUR{UziAITMEE^Dk:U]aX*iytucw9RW$SU-OMo#S<cD"i.DCY4MwCрY% O٘v^%`8|q%Y-` yWރIR&ُRs	X?>?I~v&9r{ۿZjjji#L`   0  1=mR`00eT30K8>>;̧t!00?ZL*Xp쫪2># 0 ##""# 20?803:"3< ?# @T8"#  !4 p: 0  3 <; LË1<>[#4V<V<S<4>23/n  8 p :ȇO+c뮪Ӿ\
(.jTZfѺUi6냖T5"j{fVڕva&f=]PQm&a]?ESzj*Vتy=Kd`FO#8ᅷg)е6%vTX]֭bqV]ZWLֵ%PdXRUe'YVA%TUiITIIRUoVYI[RU'߸+MOF=:!΃%d⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ.	Gq4]D&$0tJR>^py&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%zJ}c71+odȑgۃ#a1FY_OOש斊<9RJJY*3\hCl-*	;¿\mCآ5U0/lIJ4m킘46Z-o%긇{_RH2pgdzHW+{+7nL]V<TwbiM
ha0gfbjEVWV@+ݫU  '<AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>FivePrimePhosphorylated</UpstreamModification>
<DownstreamModification>FivePrimePhosphorylated</DownstreamModification>
</AdditionalSequenceProperties>

  <?xml version="1.0"?><Features nextValidID="15"><Feature recentID="14" name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="7890" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="7099-7890" color="#ccffcc" type="standard" translated="1"/><Segment name="signal sequence" range="7891-7959" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6340-6928" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3081-3460" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3463-3738" color="#ffffff" type="standard"/></Feature><Feature recentID="9" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7960-8064" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4890-4945" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment name="-10" range="5986-5992" color="#ffffff" type="standard"/><Segment range="5993-6010" color="#ffffff" type="standard"/><Segment name="-35" range="6011-6016" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6031-6052" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5899-5917" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5846-5862" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5938-5954" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5962-5978" color="#31849b" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
   <?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  G<Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pT2K-CAGGS</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Created>2014.3.19</Created>
<LastModified>2014.3.19</LastModified>
<CreatedBy>Trial User</CreatedBy>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
  ,     m  i         iG59:R\6YxEPDGM`=]n Ҿ?ôQt`ь/4F*P~y}m+:θ;YTO3=/dx?&YS*6lᾀ@?~j+|<%8=0Ȩb>2y)9y;0 xq+G/ʨA@Kof 	q]+.:<̺.88l"xZ@x~Q},$bQ)`,<pߩ|G)F.~́/!>?S>.ϱC<pomk\<PʋjhX_q6@.B⅋!AM+:Q.$0_~t#0S܉g~5C#ܢՃLP'`v"
U\*Ue`A8#,NrWu+L1NJۍS#HO9vqKcCAy3!Jw!|lzpo7S'^urmSE^up#Nywl18uMQ\E3v+!u%L<ǹCcS	<9r_	yc5jTeQjKP XT߉
S%jC`UYJѵ
AtDtff	M+;@FJ}b@XjҳθD3G'&ҍz 8.L ]*R]$ŚR[i=[ݞVJ`thhZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%zJ}c71+odȑgۃ#a1FY_OOש斊<9RJJY*3\hCl-*	;¿\mCآ5U0/lIJ4m킘46Z-o%긇{_RH2pgdzHW+{+7nL]V<TwbiM
ha0gfbjEVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxDcgьV|C=8xl#sӃ 5iJ\
rW3PX#cPFt O nbM|߈3$CK֢m YӅ>dIFUF.U@  '<AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>FivePrimePhosphorylated</UpstreamModification>
<DownstreamModification>FivePrimePhosphorylated</DownstreamModification>
</AdditionalSequenceProperties>

  <?xml version="1.0"?><Features nextValidID="81"><Feature recentID="80" name="mscv 5' primer" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1333-1355" color="#0080ff" type="standard"/><Q name="note"><V text="mscv 5' primer"/></Q></Feature><Feature recentID="79" name="SJVL-977_Max=753" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6056-6075" color="#00ff00" type="standard"/><Q name="note"><V text="RSET Sequencing.  Max recorded sequencing read =  753 bases.  Perfect match Tm = 56.3 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CGGGCCTCTT CGCTATTACG .  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="78" name="SJVL-779_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6445-6467" color="#00ff00" type="standard"/><Q name="note"><V text="MLV reading right into LTR.  Max recorded  sequencing read =  bases.  Perfect match Tm = 44.8 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is ATAGAGTACG AGCCATAGAT AAA.  This primer  matches this site perfectly."/></Q></Feature><Feature recentID="77" name="SJVL-778_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6390-6411" color="#00ff00" type="standard"/><Q name="note"><V text="MLV reading right into LTR.  Max recorded  sequencing read =  bases.  Perfect match Tm = 50.1 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GATATCAGTG GTCCAGGCTC TA.  This primer  matches this site perfectly."/></Q></Feature><Feature recentID="76" name="SJVL-344_Max=142" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6216-6242" color="#00ff00" type="standard"/><Q name="note"><V text="AAV 5 vector sequencing.  Max recorded sequencing  read = 142 bases.  Perfect match Tm = 73.2 C, although  for oligos less than 35 bp, only the 3 prime 17 nt must  match to trigger annotation.  The actual sequence of the  primer is GCCTGCCACC ATACCCACGC CGAAACA.  This primer  matches this site perfectly."/></Q></Feature><Feature recentID="75" name="SJVL-21_Max=932" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6123-6146" color="#00ff00" type="standard"/><Q name="note"><V text="Cloning vector sequencing primer, was HGTI-3334.   Max recorded sequencing read = 932 bases.  Perfect match  Tm = 66.1 C, although for oligos less than 35 bp, only  the 3 prime 17 nt must match to trigger annotation.  The  actual sequence of the primer is cgccagggtt ttcccagtca  cgac.  This primer matches this site perfectly."/></Q></Feature><Feature recentID="74" name="SJVL-12_Max=851" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6450-6479" color="#00ff00" type="standard"/><Q name="note"><V text="MLV sequencing.  Was HGTI-1627.  Max recorded  sequencing read = 851 bases.  Perfect match Tm = 54.4 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GTACGAGCCA TAGATAAAAT AAAAGATTTT .  This  primer matches this site perfectly."/></Q></Feature><Feature recentID="73" name="SJVL-8_Max=820" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="6406-6431" color="#00ff00" type="standard"/><Q name="note"><V text="MLV sequecing into genes, downstream.  Was  HGTI-1293, or JSL-502.  Max recorded sequencing read =  820 bases.  Perfect match Tm = 51.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is tgatattgtt gagtcaaaac tagagc.  This primer matches  this site completely."/></Q></Feature><Feature recentID="72" name="MLV_env_pPu" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6364-6505" color="#00ffff" type="standard"/><Q name="note"><V text="142 bp upstream of MoMLV right LTR, as junk in  MSCV."/></Q></Feature><Feature recentID="71" name="SJVL-717_Max=780" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="5685-5719" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing primer in pBR322, reading towards the  EcoR I site.  Max recorded sequencing read = 780 bases.   Perfect match Tm = 71.9 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  CCCGAAAAGT GCCACCTGAC GTCTAAGAAA CCATT.  This primer  matches this site perfectly."/></Q></Feature><Feature recentID="70" name="SJVL-120_Max=770" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="5879-5904" color="#00ff00" type="standard"/><Q name="note"><V text="AAV-CAGG sequencing primers for TRs.  Max recorded  sequencing read = 770 bases.  Perfect match Tm = 76.6 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GCGTCAGCGG GTGTTGGCGG GTGTCG.  This  primer matches this site perfectly."/></Q></Feature><Feature recentID="69" name="Plasmid_Frag" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="5771-5996" color="#00ffff" type="standard"/><Q name="note"><V text="Fragment of bacterial plasmid used in some cloning  vectors.  Loose match to the ColE1 plasmid NC_001371,  1381..1647.  Not sure if functionally significant."/></Q></Feature><Feature recentID="68" name="Tn3_frag" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="4564-5770" color="#00ffff" type="standard"/><Q name="note"><V text="Piece of Tn3 found in some cloning vectors, which  carries beta lactamase gene.  3751..4957 of V00613.1."/></Q></Feature><Feature recentID="67" name="bla" directionality="2" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4714-5571" color="#ffff00" type="standard"/><Q name="note"><V text="B-lactamase, confers ampicillin resistance, no Pst  I."/></Q></Feature><Feature recentID="66" name="SJVL-788_Max=223" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="4071-4092" color="#00ff00" type="standard"/><Q name="note"><V text="plasmid backbone primer for sequencing  scAAVv2-GFP.  Max recorded sequencing read = 223 bases.   Perfect match Tm = 62.9 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  GGCAGGGTCG GAACAGGAGA GC.  This primer matches this site  completely."/></Q></Feature><Feature recentID="65" name="SJVL-787_Max=527" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="4033-4053" color="#00ff00" type="standard"/><Q name="note"><V text="plasmid backbone primer for sequencing  scAAVv2-GFP.  Max recorded sequencing read = 527 bases.   Perfect match Tm = 57.9 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  GGGGGAAACG CCTGGTATCT T.  This primer matches this site  completely."/></Q></Feature><Feature recentID="64" name="High_copy" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="4372-4415" color="#00ffff" type="standard"/><Q name="note"><V text="Tag for High copy plasmid replication xxpUC18xx"/></Q></Feature><Feature recentID="63" name="SJVL-343_Max=121" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3617-3647" color="#00ff00" type="standard"/><Q name="note"><V text="AAV 5 vector sequencing.  Max recorded sequencing  read = 121 bases.  Perfect match Tm = 64.1 C, although  for oligos less than 35 bp, only the 3 prime 17 nt must  match to trigger annotation.  The actual sequence of the  primer is ATAAAGTGTA AAGCCTGGGG TGCCTAATGA G.  This  primer matches this site perfectly."/></Q></Feature><Feature recentID="62" name="SJVL-17_Max=779" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3715-3737" color="#00ff00" type="standard"/><Q name="note"><V text="Cloning vector sequencing primer.  Was HGTI-1880.   Max recorded sequencing read = 779 bases.  Perfect match  Tm = 59.4 C, although for oligos less than 35 bp, only  the 3 prime 17 nt must match to trigger annotation.  The  actual sequence of the primer is gttggccgat tcattaatgc  agc.  This primer matches this site completely."/></Q></Feature><Feature recentID="61" name="ori" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="3765-4563" color="#00ffff" type="standard"/><Q name="note"><V text="Origin of replication for some high copy bacterial  plasmids.  Loose match to ColE1 plasmid sequence  NC_001371, from 582..1381, which contains the RNA II  primer transcript.  Critical t mutation defining high  copy replication is in lower case."/></Q></Feature><Feature recentID="60" name="lacO" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3574-3590" color="#00ffff" type="standard"/><Q name="note"><V text="lac operator; bound by LacI repressor to repress  lac operon expression"/></Q></Feature><Feature recentID="59" name="SJVL-903_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3247-3266" color="#00ff00" type="standard"/><Q name="note"><V text="Biotinylated oligo for MLV vector DNA  purification.  Max recorded sequencing read =  bases.   Perfect match Tm = 58.5 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  AGCCCACAAC CCCTCACTCG .  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="58" name="SJVL-783_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3218-3234" color="#00ff00" type="standard"/><Q name="note"><V text="MLV ECT3- U3 primer facing right.  Max recorded  sequencing read =  bases.  Perfect match Tm = 57.0 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GCGCTTCTGC TCCCCGA.  This primer matches  this site perfectly."/></Q></Feature><Feature recentID="57" name="SJVL-781_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3226-3248" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U3 facing right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 57.4 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCTCCCCGAG CTCAATAAAA GAG.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="56" name="SJVL-780_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3244-3264" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U3, same as 725 but shifted left a little.   Max recorded sequencing read =  bases.  Perfect match Tm  = 56.3 C, although for oligos less than 35 bp, only the 3  prime 17 nt must match to trigger annotation.  The actual  sequence of the primer is AAGAGCCCAC AACCCCTCAC T.  This  primer matches this site perfectly."/></Q></Feature><Feature recentID="55" name="SJVL-770_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3368-3393" color="#00ff00" type="standard"/><Q name="note"><V text="More MLV LTR primers.  Max recorded sequencing  read =  bases.  Perfect match Tm = 57.4 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is GGGTAGTCAA TCACTCAGAG GAGACC.  This primer matches  this site completely."/></Q></Feature><Feature recentID="54" name="SJVL-769_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3323-3344" color="#00ff00" type="standard"/><Q name="note"><V text="More MLV LTR primers.  Max recorded sequencing  read =  bases.  Perfect match Tm = 55.9 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is AAGTCGGATG CAACTGCAAG AG.  This primer matches this  site completely."/></Q></Feature><Feature recentID="53" name="SJVL-768_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3306-3330" color="#00ff00" type="standard"/><Q name="note"><V text="More MLV LTR primers.  Max recorded sequencing  read =  bases.  Perfect match Tm = 58.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCAAGAGGG TTTATTGGAT ACACG.  This primer matches this  site completely."/></Q></Feature><Feature recentID="52" name="SJVL-751_Max=580" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3346-3365" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U5 facing left, pcr or sequencing.  Max  recorded sequencing read = 580 bases.  Perfect match Tm =  57.3 C, although for oligos less than 35 bp, only the 3  prime 17 nt must match to trigger annotation.  The actual  sequence of the primer is CCCAAGGAAC AGCGAGACCA .  This  primer matches this site completely."/></Q></Feature><Feature recentID="51" name="SJVL-746_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3368-3391" color="#00ff00" type="standard"/><Q name="note"><V text="ABI designed B2 PCR left.  Max recorded sequencing  read =  bases.  Perfect match Tm = 50.5 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is GGTCTCCTCT GAGTGATTGA CTAC.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="50" name="SJVL-740_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3253-3282" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U3-R junction mutagenesis plasmid.  Creates  Asc I site.  Max recorded sequencing read =  bases.   Perfect match Tm = 80.2 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  CAACCCCTCA CTCGGCGCGC CAGTCCTCCG .  This primer matches  this site perfectly."/></Q></Feature><Feature recentID="49" name="SJVL-728_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3194-3216" color="#00ff00" type="standard"/><Q name="note"><V text="mlv pbs primer  Max recorded sequencing read =   bases.  Perfect match Tm = 58.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  AGTTCGCTTC TCGCTTCTGT TCG.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="48" name="SJVL-726_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3328-3349" color="#00ff00" type="standard"/><Q name="note"><V text="MLV pbs pcr  Max recorded sequencing read =   bases.  Perfect match Tm = 58.6 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCAGTTGCAT CCGACTTGTG GT.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="47" name="SJVL-725_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3248-3266" color="#00ff00" type="standard"/><Q name="note"><V text="MLV PBS pcr  Max recorded sequencing read =   bases.  Perfect match Tm = 57.4 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCCCACAACC CCTCACTCG.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="46" name="SJVL-376_Max=494" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3187-3211" color="#00ff00" type="standard"/><Q name="note"><V text="In MSCV U3, facing left.  Max recorded sequencing  read = 494 bases.  Perfect match Tm = 60.6 C, although  for oligos less than 35 bp, only the 3 prime 17 nt must  match to trigger annotation.  The actual sequence of the  primer is AGAAGCGAGA AGCGAACTGA TTGGT.  This primer  matches this site completely."/></Q></Feature><Feature recentID="45" name="MLV_LTR_tip" directionality="2" type="repeat_region" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3401-3411" color="#000000" type="standard"/><Q name="note"><V text="Tip of both MLV and MPSV LTR. "/></Q></Feature><Feature recentID="44" name="muGenomic" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3412-3567" color="#00ffff" type="standard"/><Q name="note"><V text="Murine genomic DNA flanking Abelson murine  leukemia virus integrated genome, as described in  V01541.1"/></Q></Feature><Feature recentID="43" name="R_MSCV" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3269-3336" color="#000000" type="standard"/><Q name="note"><V text="The right R region from MSCV vectors, which is  left half MPSV, and right half MLV, as if they used the  Kpn I site."/></Q></Feature><Feature recentID="42" name="U5_MLV" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="3337-3411" color="#000000" type="standard"/><Q name="note"><V text="U5 region of Moloney MLV."/></Q></Feature><Feature recentID="41" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="2712-2744" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="40" name="SJVL-498_Max=860" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="2683-2701" color="#00ff00" type="standard"/><Q name="note"><V text="GFP PCR right.  Max recorded sequencing read = 860  bases.  Perfect match Tm = 63.2 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  CGGCGGCGGT CACGAACTC.  This primer matches this site  completely."/></Q></Feature><Feature recentID="39" name="SJVL-403_Max=772" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="2561-2589" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing Primers for HRST.  Max recorded  sequencing read = 772 bases.  Perfect match Tm = 71.8 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GTCGCCGATG GGGGTGTTCT GCTGGTAGT.  This  primer matches this site completely."/></Q></Feature><Feature recentID="38" name="SJVL-12_Max=851" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2843-2872" color="#00ff00" type="standard"/><Q name="note"><V text="MLV sequencing.  Was HGTI-1627.  Max recorded  sequencing read = 851 bases.  Perfect match Tm = 54.4 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GTACGAGCCA TAGATAAAAT AAAAGATTTT .  This  primer does not match this site perfectly."/></Q></Feature><Feature recentID="37" name="LTR" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2899-3411" color="#000000" type="standard"/><Q name="note"><V text="MSCV left LTR, which is purely PCMV derived."/></Q></Feature><Feature recentID="36" name="MSCV_pPu" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2854-2898" color="#00ffff" type="standard"/><Q name="note"><V text="Extent of polypurine tract in MSCV vectors.  1st  base is unique-ifier.  Also occurs upstream of left  LTR."/></Q></Feature><Feature recentID="35" name="AAV5_TR_C" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2586-2593" color="#000000" type="standard"/><Q name="note"><V text="AAV serotype 5 ITR region C."/></Q></Feature><Feature recentID="34" name="U3_PCMV" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2899-3268" color="#000000" type="standard"/><Q name="note"><V text="U3 region from PCMV virus, which was published as  modified by passaging to enable more efficient infection  of embryonal carcinoma cells.  It has lost one of the 75  bp repeats, but other than that is nearly identical to  MPSV xxwhen you take into account the errors in the  genbank sequencexx."/></Q></Feature><Feature recentID="33" name="MLV_LTR_tip" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2899-2909" color="#000000" type="standard"/><Q name="note"><V text="Tip of both MLV and MPSV LTR. "/></Q></Feature><Feature recentID="32" name="SJVL-497_Max=794" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2416-2439" color="#00ff00" type="standard"/><Q name="note"><V text="GFP PCR left.  Max recorded sequencing read = 794  bases.  Perfect match Tm = 67 C, although for oligos less  than 35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  GACGGCAACA TCCTGGGGCA CAAG.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="31" name="SJVL-404_Max=836" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="2096-2121" color="#00ff00" type="standard"/><Q name="note"><V text="Sequencing Primers for HRST.  Max recorded  sequencing read = 836 bases.  Perfect match Tm = 73.4 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is CTCGCCCTCG CCGGACACGC TGAACT.  This  primer matches this site completely."/></Q></Feature><Feature recentID="30" name="GFP" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2017-2733" color="#00ff00" type="standard"/><Q name="note"><V text="EGFP from Clontech pEGFP-N1 and C1, such that both  polylinkers are excluded."/></Q></Feature><Feature recentID="29" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1875-1896" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="28" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="1491-1510" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="27" name="SJVL-358_Max=772" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1364-1392" color="#00ff00" type="standard"/><Q name="note"><V text="MSCV pgk neo sequencing.  Max recorded sequencing  read = 772 bases.  Perfect match Tm = 65.6 C, although  for oligos less than 35 bp, only the 3 prime 17 nt must  match to trigger annotation.  The actual sequence of the  primer is ATCCTCCCTT TATCCAGCCC TCACTCCTT.  This primer  matches this site perfectly."/></Q></Feature><Feature recentID="26" name="SJVL-4_Max=816" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="1020-1048" color="#00ff00" type="standard"/><Q name="note"><V text="MLV sequencing.  Was HGTI-1067.  Max recorded  sequencing read = 816 bases.  Perfect match Tm = 65.4 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is ATCCGCTCGA CATCTTTCCA GTGACCTAA.  This  primer matches this site completely."/></Q></Feature><Feature recentID="25" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1455-2016" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature recentID="24" name="SJVL-711_Max=870" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="677-700" color="#00ff00" type="standard"/><Q name="note"><V text="MLV sequencing primer.  In Psi facing left.  Max  recorded sequencing read = 870 bases.  Perfect match Tm =  66.3 C, although for oligos less than 35 bp, only the 3  prime 17 nt must match to trigger annotation.  The actual  sequence of the primer is CCACGGGTCC GCCAGATACA GAGC.   This primer matches this site completely."/></Q></Feature><Feature recentID="23" name="SJVL-629_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="882-900" color="#00ff00" type="standard"/><Q name="note"><V text="MSCV csgag real time primer right.  Max recorded  sequencing read =  bases.  Perfect match Tm = 52.6 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is TTCGGTTCCA AACCGAAAG.  This primer  matches this site completely."/></Q></Feature><Feature recentID="22" name="SJVL-576_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="855-880" color="#00ff00" type="standard"/><Q name="note"><V text="MLV real time probe.  May work on MSCV.  Max  recorded sequencing read =  bases.  Perfect match Tm =  64.9 C, although for oligos less than 35 bp, only the 3  prime 17 nt must match to trigger annotation.  The actual  sequence of the primer is CAGTTCCCGC CTCCGTCTGA ATTTTT.   This primer matches this site perfectly."/></Q></Feature><Feature recentID="21" name="SJVL-574_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="829-853" color="#00ff00" type="standard"/><Q name="note"><V text="MLV csgag real time primer left.  May work on  MSCV.  Max recorded sequencing read =  bases.  Perfect  match Tm = 54.7 C, although for oligos less than 35 bp,  only the 3 prime 17 nt must match to trigger annotation.   The actual sequence of the primer is TTCTGGTAGG  AGACGAGAAC CTAAA.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="20" name="SJVL-496_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="716-739" color="#00ff00" type="standard"/><Q name="note"><V text="MSCV Psi PCR right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 72 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  CCAGGGTTGC GGCCGGGTGT TCAG.  This primer matches this  site completely."/></Q></Feature><Feature recentID="19" name="SJVL-495_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="582-603" color="#00ff00" type="standard"/><Q name="note"><V text="MSCV Psi PCR left.  Max recorded sequencing read  =  bases.  Perfect match Tm = 64.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  CTGGCCAGCG GTCGTTTCGT GT.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="18" name="SJVL-9_Max=823" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1000-1029" color="#00ff00" type="standard"/><Q name="note"><V text="MLV sequencing.  Was HGTI-1606.  Max recorded  sequencing read = 823 bases.  Perfect match Tm = 61.3 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is CCACTCCCTT AAGTTTGACC TTAGGTCACT .  This  primer matches this site perfectly."/></Q></Feature><Feature recentID="17" name="PBS-Q" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="516-533" color="#00ff00" type="standard"/><Q name="note"><V text="tRNA binding site for MMP vectors, for Glutamine  tRNA."/></Q></Feature><Feature recentID="16" name="MLV_LTR_tip" directionality="2" type="repeat_region" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="503-513" color="#000000" type="standard"/><Q name="note"><V text="Tip of both MLV and MPSV LTR. "/></Q></Feature><Feature recentID="15" name="Psi_MSCV" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="534-1403" color="#00ffff" type="standard"/><Q name="note"><V text="Everything from PBS-Q to end of MLV sequence at  Psi position in MSCV vectors.  Everything left of Pst I  site is from endogenous murine PBS-Q virus.  After that,  but for ATG -&amp;gt; AG mutation in gag, it is identical to  moloney MLV.  This version does not have an Avr II site,  and is confirmed in the MSCV-neo construct."/></Q></Feature><Feature recentID="14" name="MA_p15" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="986-1376" color="#ffff00" type="standard"/><Q name="note"><V text="MoMLV matrix protein from gag, without initiator  methionine.  As found in MSCV vectors.  In wild type  virus, ATG proceeds this sequence."/></Q></Feature><Feature recentID="13" name="SJVL-903_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="349-368" color="#00ff00" type="standard"/><Q name="note"><V text="Biotinylated oligo for MLV vector DNA  purification.  Max recorded sequencing read =  bases.   Perfect match Tm = 58.5 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  AGCCCACAAC CCCTCACTCG .  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="12" name="SJVL-783_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="320-336" color="#00ff00" type="standard"/><Q name="note"><V text="MLV ECT3- U3 primer facing right.  Max recorded  sequencing read =  bases.  Perfect match Tm = 57.0 C,  although for oligos less than 35 bp, only the 3 prime 17  nt must match to trigger annotation.  The actual sequence  of the primer is GCGCTTCTGC TCCCCGA.  This primer matches  this site perfectly."/></Q></Feature><Feature recentID="11" name="SJVL-781_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="328-350" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U3 facing right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 57.4 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCTCCCCGAG CTCAATAAAA GAG.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="10" name="SJVL-780_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="346-366" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U3, same as 725 but shifted left a little.   Max recorded sequencing read =  bases.  Perfect match Tm  = 56.3 C, although for oligos less than 35 bp, only the 3  prime 17 nt must match to trigger annotation.  The actual  sequence of the primer is AAGAGCCCAC AACCCCTCAC T.  This  primer matches this site perfectly."/></Q></Feature><Feature recentID="9" name="SJVL-740_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="355-384" color="#00ff00" type="standard"/><Q name="note"><V text="MLV U3-R junction mutagenesis plasmid.  Creates  Asc I site.  Max recorded sequencing read =  bases.   Perfect match Tm = 80.2 C, although for oligos less than  35 bp, only the 3 prime 17 nt must match to trigger  annotation.  The actual sequence of the primer is  CAACCCCTCA CTCGGCGCGC CAGTCCTCCG .  This primer matches  this site perfectly."/></Q></Feature><Feature recentID="8" name="SJVL-728_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="296-318" color="#00ff00" type="standard"/><Q name="note"><V text="mlv pbs primer  Max recorded sequencing read =   bases.  Perfect match Tm = 58.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  AGTTCGCTTC TCGCTTCTGT TCG.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="7" name="SJVL-725_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="350-368" color="#00ff00" type="standard"/><Q name="note"><V text="MLV PBS pcr  Max recorded sequencing read =   bases.  Perfect match Tm = 57.4 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCCCACAACC CCTCACTCG.  This primer matches this site  perfectly."/></Q></Feature><Feature recentID="6" name="SJVL-376_Max=494" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="289-313" color="#00ff00" type="standard"/><Q name="note"><V text="In MSCV U3, facing left.  Max recorded sequencing  read = 494 bases.  Perfect match Tm = 60.6 C, although  for oligos less than 35 bp, only the 3 prime 17 nt must  match to trigger annotation.  The actual sequence of the  primer is AGAAGCGAGA AGCGAACTGA TTGGT.  This primer  matches this site completely."/></Q></Feature><Feature recentID="5" name="LTR" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1-513" color="#000000" type="standard"/><Q name="note"><V text="MSCV left LTR, which is a hybrid of PCMV and a  mouse endogenous virus."/></Q></Feature><Feature recentID="4" name="dl587rev" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="407-923" color="#00ffff" type="standard"/><Q name="note"><V text="Fragment of recombinant virus isolated by  Colicelli &amp;amp; Goff xxJVirol 1986xx.  This portion of that  virus is derived from endogenous murine genomic  sequences.  It introduces several sequence variations  from MSCV, including glutamine tRNA binding site."/></Q></Feature><Feature recentID="3" name="U3_PCMV" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-370" color="#000000" type="standard"/><Q name="note"><V text="U3 region from PCMV virus, which was published as  modified by passaging to enable more efficient infection  of embryonal carcinoma cells.  It has lost one of the 75  bp repeats, but other than that is nearly identical to  MPSV xxwhen you take into account the errors in the  genbank sequencexx."/></Q></Feature><Feature recentID="2" name="MLV_LTR_tip" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1-11" color="#000000" type="standard"/><Q name="note"><V text="Tip of both MLV and MPSV LTR. "/></Q></Feature><Feature recentID="1" name="U5_PBSQ" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="440-513" color="#000000" type="standard"/><Q name="note"><V text="U5 from MFG-SxxMPSV-PBSQxx.  Derives from an  endogenous murine virus which contained the PBS-Q."/></Q></Feature><Feature recentID="0" name="R_MoPBSQ" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="371-439" color="#000000" type="standard"/><Q name="note"><V text="R region from MFG-SxxMPSVxxPBSQxx.  After the Kpn  I site, it is an endogenous murine virus.  Before that,  it is MPSV."/></Q></Feature></Features>
   <?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
   <Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created>2014.3.27</Created>
<LastModified>2014.3.27</LastModified>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
       L           Ymb@٨ٜgPij%wTfuYrEIe$ҩVUَ%TfnpVwUYk~U[YzmeZUa5YU\I{2YdSN[jMՊ}8^T.ׇvV(߆U(buk]9u:Y]jvbӎNU孖.c}ff֮(n:N(Uj)YkNaiwW5[VeNn"ke.z2]]ej
ҽ]5m땕jRgk]שKZ(
֪Z*UJ!R%ںd劦U4ifE:&8Kmiٽ]V뮤֮oJTfa1m[z)zdOcʤjjZ⡫ۖaتh[MkeYbf0cAkvV*ۭXYrܖj	Z+^3YjnꨜNEdԥYC^T|KZNBTh,q  <AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>Unmodified</UpstreamModification>
<DownstreamModification>Unmodified</DownstreamModification>
</AdditionalSequenceProperties>

   <?xml version="1.0"?><Features nextValidID="2"><Feature recentID="1" name="U6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="16-256" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="label"><V text="U6 Promoter"/></Q></Feature><Feature recentID="0" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2146-2221" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  =<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"><BindingSite location="0-44" boundStrand="0" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="65"><Component hybridizedRange="0-44" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite><BindingSite simplified="1" location="0-44" boundStrand="0" annealedBases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="65"><Component hybridizedRange="0-44" bases="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite></Primer><Primer recentID="1" name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"><BindingSite location="2237-2279" boundStrand="1" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="72"><Component hybridizedRange="2237-2279" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite><BindingSite simplified="1" location="2237-2279" boundStrand="1" annealedBases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="72"><Component hybridizedRange="2237-2279" bases="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite></Primer></Primers>
   <Notes>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created>2015.10.20</Created>
<LastModified>2015.10.20</LastModified>
<CreatedBy>Tumor Cell Biology</CreatedBy>
<TransformedInto>Unspecified</TransformedInto>
</Notes>
       H           ,qkffqCDFZExQIg!%JZbVf:|RjƮ TW|XMVwcfaVneIj+WYUeUs%*ߩeފL-;m6V+ySh^ۭX~Vծ[ӭtj)euYڗ]oN::.WZ}Z}WX867AW]v>eY:+Y^(mZ֎ݖ:M*Xnfj*JfZ^ʡtk}wU+J1tնVUIK6ilbKv^/MjXjX+ZjV*THcjVV땒*V9n-f^SUuZ򛮺Zڕ)Ru^jƹmUY<jS*e֕h/n[Ubn-6-Yfm}jծjuکXnccefkrY%jJixUeN;r*9ReKmzR,#k99PH,  '<AdditionalSequenceProperties>
<UpstreamStickiness>0</UpstreamStickiness>
<DownstreamStickiness>0</DownstreamStickiness>
<UpstreamModification>FivePrimePhosphorylated</UpstreamModification>
<DownstreamModification>FivePrimePhosphorylated</DownstreamModification>
</AdditionalSequenceProperties>

  <?xml version="1.0"?><Features nextValidID="2"><Feature recentID="1" name="U6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7-247" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="label"><V text="U6 Promoter"/></Q></Feature><Feature recentID="0" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2137-2212" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"><BindingSite location="2228-2261" boundStrand="1" annealedBases="GTCGACCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="60"><Component bases="ATGGGCCCTC"/><Component hybridizedRange="2255-2261" bases="GTCGACC"/><Component hybridizedRange="2228-2253" bases="AATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite><BindingSite simplified="1" location="2228-2255" boundStrand="1" annealedBases="CCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="60"><Component bases="ATGGGCCCTCGTCGA"/><Component hybridizedRange="2228-2255" bases="CCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite></Primer><Primer recentID="0" name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"><BindingSite location="0-35" boundStrand="0" annealedBases="GTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="60"><Component bases="AATAATCAAT"/><Component hybridizedRange="0-4" bases="GTCGA"/><Component hybridizedRange="6-35" bases="GAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite><BindingSite simplified="1" location="6-35" boundStrand="0" annealedBases="GAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="60"><Component bases="AATAATCAATGTCGA"/><Component hybridizedRange="6-35" bases="GAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite></Primer></Primers>
   <Notes>
<Type>Natural</Type>
<Created>2015.10.9</Created>
<LastModified>2015.10.20</LastModified>
<CreatedBy>Tumor Cell Biology</CreatedBy>
<SequenceClass>UNA</SequenceClass>
</Notes>
  <?xml version="1.0"?><HistoryTree><Node name="pXPR_003-puro-IRES-CFP[29997].dna" type="DNA" seqLen="11489" ID="10" circular="1" operation="inFusionCloning"><RegeneratedSite name="MluI" pos="3184" siteCount="1"/><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="1882..3183" colors="red"/></TopStrand><BottomStrand><ColorRange range="1882..3183" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" name1="MluI" val1="1878" siteCount1="1"/><InputSummary manipulation="overlapAndInsert" val1="0" val2="1331"/><Node name="pXPR_003 withStuffer[1].dna" type="DNA" seqLen="10183" ID="0" circular="1" resurrectable="1" operation="invalid"><Features><Feature name="Filler" type="misc_binding" allowSegmentOverlaps="0"><Segment range="7998-9882" color="#a6acb3" type="standard"/><Q name="note"><V text="Filler"/></Q></Feature><Feature name="EF1a" type="promoter" allowSegmentOverlaps="0"><Segment range="7-1265" color="#ffffff" type="standard"/><Q name="note"><V text="EF1a"/></Q></Feature><Feature name="AmpR" directionality="1" type="CDS" allowSegmentOverlaps="0"><Segment range="3953-4813" color="#993366" type="standard"/><Q name="note"><V text="AmpR"/></Q></Feature><Feature name="pUC Ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5074-5689" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="pUC Ori"/></Q></Feature><Feature name="Puro" directionality="1" type="CDS" allowSegmentOverlaps="0"><Segment range="1278-1877" color="#993366" type="standard"/><Q name="note"><V text="Puro"/></Q></Feature><Feature name="WPRE" type="misc_binding" allowSegmentOverlaps="0"><Segment range="1893-2481" color="#a6acb3" type="standard"/><Q name="note"><V text="WPRE"/></Q></Feature><Feature name="F1 ori" type="rep_origin" allowSegmentOverlaps="0"><Segment range="3367-3822" color="#ffff00" type="standard"/><Q name="note"><V text="F1 ori"/></Q></Feature><Feature name="5'LTR" type="misc_structure" allowSegmentOverlaps="0"><Segment range="6037-6449" color="#a6acb3" type="standard"/><Q name="note"><V text="5'LTR"/></Q></Feature><Feature name="gag" type="misc_feature" allowSegmentOverlaps="0"><Segment range="6604-6968" color="#a6acb3" type="standard"/><Q name="note"><V text="gag"/></Q></Feature><Feature name="RRE" type="misc_feature" allowSegmentOverlaps="0"><Segment range="7114-7355" color="#a6acb3" type="standard"/><Q name="note"><V text="RRE"/></Q></Feature><Feature name="3'LTR" type="3'UTR" allowSegmentOverlaps="0"><Segment range="2552-2787" color="#a6acb3" type="standard"/><Q name="note"><V text="3'LTR"/></Q></Feature><Feature name="SV40 polyadenylation signal" type="polyA_signal" allowSegmentOverlaps="0"><Segment range="2864-2994" color="#a6acb3" type="standard"/><Q name="note"><V text="SV40 polyadenylation signal"/></Q></Feature><Feature name="CPPT" type="misc_feature" allowSegmentOverlaps="0"><Segment range="10014-10131" color="#a6acb3" type="standard"/><Q name="note"><V text="CPPT"/></Q></Feature><Feature name="chRNA" type="misc_feature" allowSegmentOverlaps="0"><Segment range="9883-9964" color="#a6acb3" type="standard"/><Q name="note"><V text="chRNA"/></Q></Feature><Feature name="SV40 origin of replication" type="rep_origin" allowSegmentOverlaps="0"><Segment range="3074-3149" color="#ffff00" type="standard"/><Q name="note"><V text="SV40 origin of replication"/></Q></Feature><Feature name="T7" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="3180-3199" color="#ffffff" type="standard"/><Q name="note"><V text="T7"/></Q></Feature></Features></Node><Node name="Fragment" type="DNA" seqLen="1332" ID="9" circular="0" upstreamModification="Unmodified" downstreamModification="Unmodified" resurrectable="1" operation="amplifyFragment"><Features><Feature name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="36-597" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature name="eCFP" type="misc_feature" allowSegmentOverlaps="0"><Segment range="598-1314" color="#00ccff" type="standard"/></Feature></Features><Oligo name="Fragment.FOR" sequence="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"/><Oligo name="Fragment.REV" sequence="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"/><Parameter name="polymerase" val="polymerase that creates blunt ends"/><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="15..1316" colors="red"/></TopStrand><BottomStrand><ColorRange range="15..1316" colors="red"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="0..14" colors="red"/><ColorRange range="1317..1317" colors="red"/><ColorRange range="1321..1331" colors="red"/></TopStrand><BottomStrand><ColorRange range="0..14" colors="red"/><ColorRange range="1317..1317" colors="red"/><ColorRange range="1321..1331" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="amplify" val1="7083" val2="8388"/><Node name="pT2K CAGGS IRES CFP U6 stuffer (infusion).dna" type="DNA" seqLen="11681" ID="8" circular="1" resurrectable="1" operation="insert"><Features><Feature name="stuffer" type="misc_feature" allowSegmentOverlaps="0"><Segment range="3199-5087" color="#a6acb3" type="standard"/></Feature><Feature name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="10697-11488" color="#ccffcc" type="standard"/><Segment name="signal sequence" range="11489-11557" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature><Feature name="eCFP" type="misc_feature" allowSegmentOverlaps="0"><Segment range="7666-8382" color="#00ccff" type="standard"/></Feature><Feature name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9938-10526" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="7104-7665" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="5335-5714" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="5717-5992" color="#ffffff" type="standard"/></Feature><Feature name="U6 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5092-5332" color="#ffffff" type="standard"/><Q name="label"><V text="U6 Promoter"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="11558-11662" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0"><Segment range="8488-8543" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment name="-10" range="9584-9590" color="#ffffff" type="standard"/><Segment range="9591-9608" color="#ffffff" type="standard"/><Segment name="-35" range="9609-9614" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6958-6987" color="#a020f0" type="standard"/></Feature><Feature name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9629-9650" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9497-9515" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9444-9460" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9536-9552" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9560-9576" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="3081-3082" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><Primers><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Fragment.FOR" sequence="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"><BindingSite simplified="1" location="7083-7102" boundStrand="0" annealedBases="CTCGAGCGGGATCAATTCCG" meltingTemperature="60"><Component bases="CGGTGCCTGAACGCG"/><Component hybridizedRange="7083-7102" bases="CTCGAGCGGGATCAATTCCG"/></BindingSite></Primer><Primer recentID="1" name="Fragment.REV" sequence="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"><BindingSite simplified="1" location="8363-8388" boundStrand="1" annealedBases="CGCTTACTTGTACAGCTCGTCCATG" meltingTemperature="59"><Component bases="TGTCGACTTAA"/><Component hybridizedRange="8386-8388" bases="CGC"/><Component bases="G"/><Component hybridizedRange="8363-8384" bases="TTACTTGTACAGCTCGTCCATG"/></BindingSite></Primer></Primers><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7083..8388" colors="black"/><ColorRange range="8389..7082" colors="white"/><ColorRange range="8385..8385" colors="white"/></TopStrand><BottomStrand><ColorRange range="7083..8388" colors="black"/><ColorRange range="8389..7082" colors="white"/><ColorRange range="8385..8385" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="7665..8381" colors="red"/></TopStrand><BottomStrand><ColorRange range="7665..8381" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="7665"/><Node name="pT2K CAGGS IRES CFP U6 stuffer (infusion).dna" type="DNA" seqLen="10964" ID="7" circular="1" resurrectable="1" operation="remove"><Features><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="3081-3082" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8843-8859" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8819-8835" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8727-8743" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8780-8798" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8912-8933" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6958-6987" color="#a020f0" type="standard"/></Feature><Feature name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment name="-10" range="8867-8873" color="#ffffff" type="standard"/><Segment range="8874-8891" color="#ffffff" type="standard"/><Segment name="-35" range="8892-8897" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0"><Segment range="7771-7826" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="gRNA scaffold" directionality="2" type="misc_RNA" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="3127-3202" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="10841-10945" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="U6 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5092-5332" color="#ffffff" type="standard"/><Q name="label"><V text="U6 Promoter"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="5717-5992" color="#ffffff" type="standard"/></Feature><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="5335-5714" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="7104-7665" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9221-9809" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9980-10771" color="#ccffcc" type="standard"/><Segment name="signal sequence" range="10772-10840" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="7665..9034" colors="blue-black"/><ColorRange range="6295..7664" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="7665..9034" colors="blue-black"/><ColorRange range="6295..7664" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="7665" val2="8381"/><Node name="pT2K CAGGS IRES CFP U6 stuffer (infusion).dna" type="DNA" seqLen="11681" ID="6" circular="1" resurrectable="1" operation="inFusionCloning" hideHistory="1"><Features><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="3081-3082" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9560-9576" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9536-9552" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9444-9460" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9497-9515" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9629-9650" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6958-6987" color="#a020f0" type="standard"/></Feature><Feature name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment name="-10" range="9584-9590" color="#ffffff" type="standard"/><Segment range="9591-9608" color="#ffffff" type="standard"/><Segment name="-35" range="9609-9614" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0"><Segment range="8488-8543" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="gRNA scaffold" directionality="2" type="misc_RNA" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="3127-3202" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="11558-11662" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="U6 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5092-5332" color="#ffffff" type="standard"/><Q name="label"><V text="U6 Promoter"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="5717-5992" color="#ffffff" type="standard"/></Feature><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="5335-5714" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="7104-7665" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9938-10526" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="mCherry" directionality="1" type="CDS" allowSegmentOverlaps="0"><Segment range="7666-8382" color="#ff0000" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="EGFP from Clontech pEGFP-N1 and C1, such that both  polylinkers are excluded."/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="10697-11488" color="#ccffcc" type="standard"/><Segment name="signal sequence" range="11489-11557" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature></Features><RegeneratedSite name="SalI" pos="3078" siteCount="1"/><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8382..9841" colors="blue-black"/><ColorRange range="6205..7664" colors="black-red"/><ColorRange range="7665..8381" colors="white"/></TopStrand><BottomStrand><ColorRange range="8382..9841" colors="blue-black"/><ColorRange range="6205..7664" colors="black-red"/><ColorRange range="7665..8381" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="3082..5331" colors="red"/></TopStrand><BottomStrand><ColorRange range="3082..5331" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" name1="SalI" val1="3078" siteCount1="1"/><InputSummary manipulation="flipOverlapAndInsert" val1="0" val2="2279"/><Node name="pT2K-CAGGS-IRES-mCherry.dna" type="DNA" seqLen="9427" ID="3" circular="1" resurrectable="1" operation="insertFragment" hideHistory="1"><Features><Feature name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7306-7322" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7282-7298" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7190-7206" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7243-7261" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7375-7396" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="pT2K-CAGGS insertion detection F" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="4704-4733" color="#a020f0" type="standard"/></Feature><Feature name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment name="-10" range="7330-7336" color="#ffffff" type="standard"/><Segment range="7337-7354" color="#ffffff" type="standard"/><Segment name="-35" range="7355-7360" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0"><Segment range="6234-6289" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9304-9408" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="3463-3738" color="#ffffff" type="standard"/></Feature><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="3081-3460" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="4850-5411" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7684-8272" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="mCherry" directionality="1" type="CDS" allowSegmentOverlaps="0"><Segment range="5412-6128" color="#ff0000" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="EGFP from Clontech pEGFP-N1 and C1, such that both  polylinkers are excluded."/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8443-9234" color="#ccffcc" type="standard"/><Segment name="signal sequence" range="9235-9303" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature></Features><RegeneratedSite name="EcoRI" pos="4800" siteCount="1"/><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="4800..6149" colors="red"/></TopStrand><BottomStrand><ColorRange range="4804..6153" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="replace" name1="XhoI" name2="EcoRI" val1="4806" val2="4800" siteCount1="1" siteCount2="1"/><InputSummary manipulation="insert" name1="EcoRI" name2="SalI" val1="1405" val2="2755" siteCount1="1" siteCount2="1"/><Node name="pT2K-CAGGS.dna" type="DNA" seqLen="8083" ID="1" circular="1" resurrectable="1" operation="invalid"><Features><Feature name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="7890"><Segment range="7099-7890" color="#ccffcc" type="standard"/><Segment name="signal sequence" range="7891-7959" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;β-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature><Feature name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6340-6928" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7-462" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CMV enhancer" type="enhancer" allowSegmentOverlaps="0"><Segment range="3081-3460" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human cytomegalovirus immediate early enhancer&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="chicken β-actin promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="3463-3738" color="#ffffff" type="standard"/></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7960-8064" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="β-globin poly(A) signal" type="polyA_site" allowSegmentOverlaps="0"><Segment range="4890-4945" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;rabbit&amp;nbsp;β-globin polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment name="-10" range="5986-5992" color="#ffffff" type="standard"/><Segment range="5993-6010" color="#ffffff" type="standard"/><Segment name="-35" range="6011-6016" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="CAP binding site" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6031-6052" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T7 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="630-648" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="T3 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5899-5917" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T3 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="604-620" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="SK primer" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5846-5862" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5938-5954" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5962-5978" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;lac&lt;/i&gt; repressor encoded by &lt;i&gt;lacI&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;The &lt;i&gt;lac&lt;/i&gt; repressor binds to the &lt;i&gt;lac&lt;/i&gt; operator to inhibit transcription in &lt;i&gt;E. coli&lt;/i&gt;. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="4800..4805" colors="white"/></TopStrand><BottomStrand><ColorRange range="4804..4809" colors="white"/></BottomStrand></StrandColors></HistoryColors></Node><Node name="mscv-ires-mcherry.dna" type="DNA" seqLen="6505" ID="2" circular="1" resurrectable="1" operation="invalid"><Features><Feature name="Psi_MSCV" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="534-1403" color="#00ffff" type="standard"/><Q name="note"><V text="Everything from PBS-Q to end of MLV sequence at  Psi position in MSCV vectors.  Everything left of Pst I  site is from endogenous murine PBS-Q virus.  After that,  but for ATG -&amp;gt; AG mutation in gag, it is identical to  moloney MLV.  This version does not have an Avr II site,  and is confirmed in the MSCV-neo construct."/></Q></Feature><Feature name="bla" directionality="2" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4714-5571" color="#ffff00" type="standard"/><Q name="note"><V text="B-lactamase, confers ampicillin resistance, no Pst  I."/></Q></Feature><Feature name="ori" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="3765-4563" color="#00ffff" type="standard"/><Q name="note"><V text="Origin of replication for some high copy bacterial  plasmids.  Loose match to ColE1 plasmid sequence  NC_001371, from 582..1381, which contains the RNA II  primer transcript.  Critical t mutation defining high  copy replication is in lower case."/></Q></Feature><Feature name="GFP" directionality="1" type="CDS" allowSegmentOverlaps="0"><Segment range="2017-2733" color="#00ff00" type="standard"/><Q name="note"><V text="EGFP from Clontech pEGFP-N1 and C1, such that both  polylinkers are excluded."/></Q></Feature><Feature name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0"><Segment range="1455-2016" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature name="LTR" type="repeat_region" allowSegmentOverlaps="0"><Segment range="1-513" color="#000000" type="standard"/><Q name="note"><V text="MSCV left LTR, which is a hybrid of PCMV and a  mouse endogenous virus."/></Q></Feature><Feature name="LTR" type="repeat_region" allowSegmentOverlaps="0"><Segment range="2899-3411" color="#000000" type="standard"/><Q name="note"><V text="MSCV left LTR, which is purely PCMV derived."/></Q></Feature><Feature name="PBS-Q" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="516-533" color="#00ff00" type="standard"/><Q name="note"><V text="tRNA binding site for MMP vectors, for Glutamine  tRNA."/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="1405..2754" colors="red"/><ColorRange range="0..1404" colors="white"/><ColorRange range="2755..6504" colors="white"/></TopStrand><BottomStrand><ColorRange range="1409..2758" colors="red"/><ColorRange range="0..1408" colors="white"/><ColorRange range="2759..6504" colors="white"/></BottomStrand></StrandColors></HistoryColors></Node></Node><Node name="Fragment" type="DNA" seqLen="2280" ID="5" circular="0" upstreamModification="Unmodified" downstreamModification="Unmodified" resurrectable="1" operation="amplifyFragment" hideHistory="1"><Features><Feature name="U6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="16-256" color="#ffffff" type="standard"/><Q name="label"><V text="U6 Promoter"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0"><Segment range="2146-2221" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><Oligo name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"/><Oligo name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"/><Parameter name="polymerase" val="polymerase that creates blunt ends"/><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="15..2264" colors="red"/></TopStrand><BottomStrand><ColorRange range="15..2264" colors="red"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="0..14" colors="red"/><ColorRange range="2265..2279" colors="red"/></TopStrand><BottomStrand><ColorRange range="0..14" colors="red"/><ColorRange range="2265..2279" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="amplify" val1="6" val2="2255"/><Node name="U6 stuffer.dna" type="DNA" seqLen="2262" ID="4" circular="0" upstreamModification="FivePrimePhosphorylated" downstreamModification="FivePrimePhosphorylated" resurrectable="1" operation="invalid"><Features><Feature name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0"><Segment range="2137-2212" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="U6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0"><Segment range="7-247" color="#ffffff" type="standard"/><Q name="label"><V text="U6 Promoter"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;RNA polymerase III promoter for human U6 snRNA&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><Primers><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Fragment.FOR" sequence="AATAATCAATGTCGAGAGGGCCTATTTCCCATGATTCCTTCATAT"><BindingSite simplified="1" location="6-35" boundStrand="0" annealedBases="GAGGGCCTATTTCCCATGATTCCTTCATAT" meltingTemperature="60"><Component bases="AATAATCAATGTCGA"/><Component hybridizedRange="6-35" bases="GAGGGCCTATTTCCCATGATTCCTTCATAT"/></BindingSite></Primer><Primer recentID="1" name="Fragment.REV" sequence="ATGGGCCCTCGTCGACCAATTCCCACTCCTTTCAAGACCTAGC"><BindingSite simplified="1" location="2228-2255" boundStrand="1" annealedBases="CCAATTCCCACTCCTTTCAAGACCTAGC" meltingTemperature="60"><Component bases="ATGGGCCCTCGTCGA"/><Component hybridizedRange="2228-2255" bases="CCAATTCCCACTCCTTTCAAGACCTAGC"/></BindingSite></Primer></Primers></Node></Node></Node></Node></Node></Node></Node></HistoryTree>
   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  N<?xml version="1.0"?><Features nextValidID="25"><Feature recentID="0" name="EF1a" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7-1265" color="#ffffff" type="standard"/><Q name="note"><V text="EF1a"/></Q></Feature><Feature recentID="1" name="5'UTR exon1" type="5'UTR" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="285-317" color="#a6acb3" type="standard"/><Q name="note"><V text="5'UTR exon1"/></Q></Feature><Feature recentID="2" name="Intron1" type="intron" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="318-1256" color="noColor" type="standard"/><Q name="note"><V text="Intron1"/></Q></Feature><Feature recentID="3" name="Puro" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1278-1877" color="#993366" type="standard"/><Q name="note"><V text="Puro"/></Q></Feature><Feature recentID="4" name="WPRE" type="misc_binding" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3199-3787" color="#a6acb3" type="standard"/><Q name="note"><V text="WPRE"/></Q></Feature><Feature recentID="5" name="3'LTR" type="3'UTR" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3858-4093" color="#a6acb3" type="standard"/><Q name="note"><V text="3'LTR"/></Q></Feature><Feature recentID="6" name="SV40 polyadenylation signal" type="polyA_signal" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4170-4300" color="#a6acb3" type="standard"/><Q name="note"><V text="SV40 polyadenylation signal"/></Q></Feature><Feature recentID="7" name="SV40 origin of replication" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4380-4455" color="#ffff00" type="standard"/><Q name="note"><V text="SV40 origin of replication"/></Q></Feature><Feature recentID="8" name="T7" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4486-4505" color="#ffffff" type="standard"/><Q name="note"><V text="T7"/></Q></Feature><Feature recentID="9" name="F1 ori" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4673-5128" color="#ffff00" type="standard"/><Q name="note"><V text="F1 ori"/></Q></Feature><Feature recentID="10" name="AmpR" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5259-6119" color="#993366" type="standard"/><Q name="note"><V text="AmpR"/></Q></Feature><Feature recentID="11" name="pUC Ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6380-6995" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="pUC Ori"/></Q></Feature><Feature recentID="12" name="5'LTR" type="misc_structure" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7343-7755" color="#a6acb3" type="standard"/><Q name="note"><V text="5'LTR"/></Q></Feature><Feature recentID="13" name="Psi" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7806-7943" color="#a6acb3" type="standard"/><Q name="note"><V text="Psi"/></Q></Feature><Feature recentID="14" name="gag" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7910-8274" color="#a6acb3" type="standard"/><Q name="note"><V text="gag"/></Q></Feature><Feature recentID="15" name="RRE" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8420-8661" color="#a6acb3" type="standard"/><Q name="note"><V text="RRE"/></Q></Feature><Feature recentID="16" name="hU6" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9059-9308" color="#ffffff" type="standard"/><Q name="note"><V text="hU6"/></Q></Feature><Feature recentID="17" name="Filler" type="misc_binding" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9304-11188" color="#a6acb3" type="standard"/><Q name="note"><V text="Filler"/></Q></Feature><Feature recentID="18" name="chRNA" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="11189-11270" color="#a6acb3" type="standard"/><Q name="note"><V text="chRNA"/></Q></Feature><Feature recentID="19" name="CPPT" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="11320-11437" color="#a6acb3" type="standard"/><Q name="note"><V text="CPPT"/></Q></Feature><Feature recentID="20" name="IRES" directionality="1" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1903-2464" color="#00ffff" type="standard"/><Q name="note"><V text="IRES from MSCV iresGFP.  Last 6 bp are not from  EMCV, and are unique to this vector construction."/></Q></Feature><Feature recentID="21" name="SJVL-366_Max=871" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="1939-1958" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing left.  Max recorded sequencing read  = 871 bases.  Perfect match Tm = 57.2 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is CAAACGCACA CCGGCCTTAT .  This primer matches this site  completely."/></Q></Feature><Feature recentID="22" name="SJVL-367_Max=" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="2323-2344" color="#00ff00" type="standard"/><Q name="note"><V text="in IRES facing right.  Max recorded sequencing  read =  bases.  Perfect match Tm = 59.1 C, although for  oligos less than 35 bp, only the 3 prime 17 nt must match  to trigger annotation.  The actual sequence of the primer  is TGCCCAGAAG GTACCCCATT GT.  This primer matches this  site perfectly."/></Q></Feature><Feature recentID="23" name="SJVL-749_Max=" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" visible="0"><Segment range="3182-3188" color="#00ff00" type="standard"/><Q name="note"><V text="Venus YFP pcr right.  Max recorded sequencing read  =  bases.  Perfect match Tm = 79.9 C, although for oligos  less than 35 bp, only the 3 prime 17 nt must match to  trigger annotation.  The actual sequence of the primer is  GCGGCCGCTC ACTTGTACAG CTCGTCCATG CCG.  This primer does  not match this site completely."/></Q></Feature><Feature recentID="24" name="eCFP" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2465-3181" color="#00ccff" type="standard"/></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="6"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Library Infusion R" sequence="CTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAAC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="11188-11225" boundStrand="1" annealedBases="CTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAAC" meltingTemperature="59"><Component hybridizedRange="11188-11225" bases="CTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAAC"/></BindingSite><BindingSite simplified="1" location="11188-11225" boundStrand="1" annealedBases="CTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAAC" meltingTemperature="59"><Component hybridizedRange="11188-11225" bases="CTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAAC"/></BindingSite></Primer><Primer recentID="1" name="Library Infusion  F" sequence="GGCTTTATATATCTTGTGGAAAGGACGAAACACC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9273-9306" boundStrand="0" annealedBases="GGCTTTATATATCTTGTGGAAAGGACGAAACACC" meltingTemperature="62"><Component hybridizedRange="9273-9306" bases="GGCTTTATATATCTTGTGGAAAGGACGAAACACC"/></BindingSite><BindingSite simplified="1" location="9273-9306" boundStrand="0" annealedBases="GGCTTTATATATCTTGTGGAAAGGACGAAACACC" meltingTemperature="62"><Component hybridizedRange="9273-9306" bases="GGCTTTATATATCTTGTGGAAAGGACGAAACACC"/></BindingSite></Primer><Primer recentID="2" name="Fragment.REV (2)" sequence="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"><BindingSite location="3162-3198" boundStrand="1" annealedBases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG" meltingTemperature="68"><Component hybridizedRange="3162-3198" bases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"/></BindingSite><BindingSite simplified="1" location="3162-3198" boundStrand="1" annealedBases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG" meltingTemperature="68"><Component hybridizedRange="3162-3198" bases="TGTCGACTTAACGCGTTACTTGTACAGCTCGTCCATG"/></BindingSite></Primer><Primer recentID="3" name="Fragment.FOR (2)" sequence="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"><BindingSite location="1867-1901" boundStrand="0" annealedBases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG" meltingTemperature="74"><Component hybridizedRange="1867-1901" bases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"/></BindingSite><BindingSite simplified="1" location="1867-1901" boundStrand="0" annealedBases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG" meltingTemperature="74"><Component hybridizedRange="1867-1901" bases="CGGTGCCTGAACGCGCTCGAGCGGGATCAATTCCG"/></BindingSite></Primer><Primer recentID="4" name="reverse primer for deep seq in nature paper" sequence="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="11357-11401" boundStrand="1" annealedBases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC" meltingTemperature="62"><Component hybridizedRange="11357-11401" bases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC"/></BindingSite><BindingSite simplified="1" location="11357-11401" boundStrand="1" annealedBases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC" meltingTemperature="62"><Component hybridizedRange="11357-11401" bases="CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCCC"/></BindingSite></Primer><Primer recentID="5" name="forward primer for deep seq in nature paper" sequence="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9225-9265" boundStrand="0" annealedBases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG" meltingTemperature="62"><Component hybridizedRange="9225-9265" bases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG"/></BindingSite><BindingSite simplified="1" location="9225-9265" boundStrand="0" annealedBases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG" meltingTemperature="62"><Component hybridizedRange="9225-9265" bases="AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG"/></BindingSite></Primer></Primers>
  b<Notes>
<UUID>83fcade1-5fb0-4c5b-85b4-b7a3f420d930</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="6:0:0">2018.12.26</Created>
<LastModified UTC="5:0:0">2019.5.29</LastModified>
<CreatedBy>Li Lab copy3</CreatedBy>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y      K         Unsaved Enzyme Set                                                                                                    HF                 E             Y?                                                                                                                                                                3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
  A<?xml version="1.0"?><CustomEnzymeSets><CustomEnzymeSet type="2" name="Unsaved Enzyme Set" enzymeNames="AarI Acc65I AhdI BmgBI BsaBI BsmBI BspDI BspEI BsrGI BstBI BstEII ClaI FseI KflI KpnI MluI MscI NcoI NdeI NotI NruI NsiI PflFI PmlI PpuMI RsrII ScaI SexAI SfiI SmaI SnaBI SwaI TspMI Tth111I XmaI"/></CustomEnzymeSets>
