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"'ã�×Þ­h— *oÒíw’•ÑÝÊÜë]çXŽZxJí‰ò)4LdúS  Ã!Æ!"#i#$]%o% c`¨£®9	_	k	L˜ÏÓBÛ›"¦# T€ðg	
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äð§!öø„è# Ý »Ì“@	qÚ¸©dèFuA!& <X^úÛ¼£g'%F-0�t³=f{ÆÐºñB Z � º /!ž!Ô!´$ ä F¨”û}Ž o
¸ï,?N§Ùu$†$˜$ ? Ñ þ ª
�_§æ"	È
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7U|½Ù4DÁy²'Ñ&¾tÆ$< [ ‘ZNF” ¼	õG�hé
"LßÛç¦!�›¨½ù8’¤÷£Ú1©`kLQ u x ¥ ¨ !k!ø#ÿ#$C%I% ! ñ‚?–Ü)Þz©1 “!2"û#7$Ž$M%Y%b%! p w ‡ ù 0¿7Ì3Vù,/z£$;ô
b@Êâ'#r#›#$3$Q%‚%†%%& ) ~¦›ùTÿ{®o	ô	�
%óJ§Ü;X'OQ0"s$¦$A%(  + W ž ç?ˆKVN«=
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Éêðù
0Dîsª¶ÑàâýQ‡Þk"$F$£$&< C Ö ð ®p;]  +vaá	�	bð;Õ_¤ÑÖ›!n¸œ¯ÄÒGqº§Ÿ¾å<Mp‰,†Ä?`™Ø '"##n#Y$4 3 Ä fXñpPŠ 	1	Ò	æ	\
[¶€+[šŸ¨ùòFd‹o"ðÅ¯!~51ÇŒò7z_ ‰ È L!—!Ê!$H$‚$;& » ë­iÀ€	Å	?
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G‰ìÿ%uÅ²U$ž4z’gýW € Ã ê t!å!!"$"F%è%&D  ‰AUYŽ¹p©’�ÃHö	µ	¶
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j•š:È=µÄ-Jk“2>Å6:¾A_¢6?KÝçð5V•÷aHn”¹– ¡!à!m"Š#•#»#@$Ë%Ö%5   ë ¤Ñ‹	6T�ÍôqÝ	2úY×ûX”¹ÎàjühTx)æ@°ñZoÉSàUš÷!#.#L#y#Å#ì#i$%& ÙŠÕ2©²¶ûV^xéH{A_jWØõqR}¿â–·(¥ƒ ¡ ¯!ª"®#%û%5 º2z�2ý		Á
æ jx‚ÇÜÂ»Äàjìilr”<rD|¬¶ Çí , 8 c 8!w!›!Ñ!‡"*$�%j  - 8 J âw}’%pÔßñS	¨	Ö	ƒ®Ìst€%…¬Áâñc®ºÃÉÏí )J_n}žËÑð[‘ÇúVzÉê;\Ô4mßè!W�;nw‹Ü!~–;ìå Kci½íGPc°ÈÝùt©  & < á !×!é!"Š"#h#Ì#×#& S_º
,
¢ãpAf&,cz¦ EõðA  R Z Ð �fMåŸÇ
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6T·Ø3F€x!¥ 7&jÝ#—$#   ‡U¢:Mz­<
aÈéïòø(/?¦:ÐÉW&P0 š#2$E$r$¥$L%. º Ã ï êÿo*u`h¿	œ	Ñ	å	8
ˆ$ö£øš3yÄH½ä‹ ^ ˆ Â ½!É!"#m#$X$`$·$:&4 11~ŠŽ‘Ì1õ	´	i”Uw<GwÆ@’Ö1Vºô¡Íßi>ïU`q¯ð;¶'¤+ ®!ß!ö!†"‰#Ä#)$5   7 Y ¡Þ9yð§	
Æ
¡ÇË×â2>ÀÂ�Ð+9yÓ3 •¥:ÿJOOb¯ÿ% / ; à è!™#Ö#1$q$
&D   · ¸ ¹ é/0n�)t�Ë8ï 

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 ÄÅÆ#STFv÷?ÔÕš›œ¸¹*wxxG®:‰Ša] ¬!­!¼!õ!…"!#l#ˆ#Ã#$0$&&	&  7š  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                                       A                                                                                                                                                                                  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            .   )      B   !      B   G   <      Õ   I      $      #   ,      ©   (                 A                                                                                                                                       /                         	                                                                                                                                                                                                                   
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  
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text="&lt;html&gt;&lt;body&gt;generates RNA-guided double strand breaks in DNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Cas9 (Csn1) endonuclease from the&amp;nbsp;&lt;i&gt;Streptococcus pyogenes&lt;/i&gt;&amp;nbsp;Type II CRISPR/Cas system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"/></Q></Feature><Feature recentID="8" name="EF-1Î± core promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3584-3795" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;core&amp;nbsp;promoter for human elongation factor EF-1Î±&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="P2A" directionality="1" translationMW="1987.23" type="CDS" allowSegmentOverlaps="0" cleavageArrows="8058" consecutiveTranslationNumbering="1" maxRunOn="8781" maxFusedRunOn="8781" detectionMode="exactProteinMatch"><Segment range="8005-8061" color="#cc99b2" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;2A peptide from porcine teschovirus-1 polyprotein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="ATNFSLLKQAGDVEENPGP"/></Q></Feature><Feature recentID="10" name="nucleoplasmin NLS" directionality="1" translationMW="1739.18" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="8781" maxFusedRunOn="8781" detectionMode="exactProteinMatch"><Segment range="7924-7971" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;bipartite nuclear localization signal from nucleoplasmin&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="KRPAATKKAGQAKKKK"/></Q></Feature><Feature recentID="11" name="FLAG" directionality="1" translationMW="1012.99" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="8781" maxFusedRunOn="8781" detectionMode="exactProteinMatch"><Segment range="7972-7995" color="#cc99b2" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;FLAGÂ® epitope tag, followed by an enterokinase cleavage site&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="DYKDDDDK"/></Q></Feature><Feature recentID="12" name="EGFP" directionality="1" translationMW="26942.08" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="8781" maxFusedRunOn="8781"><Segment range="8062-8778" color="#00ff00" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"/></Q></Feature><Feature recentID="13" name="gRNA scaffold" directionality="1" type="misc_RNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3447-3522" color="#00ccff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;guide RNA scaffold for the &lt;i&gt;Streptococcus pyogenes&lt;/i&gt; CRISPR/Cas9 system&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="U6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3180-3420" 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></Feature></Features>
   —<?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  §<Notes>
<UUID>fca81a24-c550-48ee-ae74-6d0715605b50</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pMXs-U6-MCS-sgRNA-Cas9-EGFP</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Created UTC="16:0:0">2016.12.22</Created>
<LastModified UTC="5:21:42">2023.11.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y      K         
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   )<?xml version="1.0"?><CustomEnzymeSets/>
