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CGWC  GWCM  WCMS  CMSC  MSCR  SCRT  CRTS  RTSC  TSCG  SCGC  jTCGR  kCGRG  lGRGR  mRGRT  nGRTY  oRTYC  pTYCW  qYCWY  rCWYC  sWYCG  tYCGK  uCGKC  vGKCY  wKCYS  xCYSS  yYSST  zSSTR  {STRS  |TRSG  }RSGK  ~SGKG  GKGC  €KGCR  �GCRG  ‚CRGC  ƒRGCC  M  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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plasmid pSG5&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLADAGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGIIGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTVSETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELAGQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLNRLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGLPGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANEARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKLAPVLQPWGQHPGDCLRVDTAVA*"/></Q></Feature><Feature recentID="2" name="ApmR" directionality="1" translationMW="28795.87" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="12888" maxFusedRunOn="12888"><Segment range="12085-12885" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aac&lt;/i&gt;(3)IV&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to apramycin (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/8892841&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/8892841&quot;&gt;Paget and Davies, 1996&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside 3-&lt;i&gt;N&lt;/i&gt;-acetyltransferase type IV &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRPLEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNVKRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLHLAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGHAFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"/></Q></Feature><Feature recentID="3" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="11293-11881" 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="4" name="traJ" directionality="1" translationMW="13464.00" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="3298-3669" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="traJ"/></Q><Q name="product"><V text="oriT-recognizing protein"/></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="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP*"/></Q></Feature><Feature recentID="5" name="oriT" type="oriT" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="3156-3265" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="6" name="gRNA scaffold" directionality="2" type="misc_RNA" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6428-6503" 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="7" name="J23119(SpeI) promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6524-6558" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial promoter (Registry of Standard Biological Parts BBa_J23119) modified to end with an SpeI site&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="lac promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment name="-10" range="10939-10945" color="#ffffff" type="standard"/><Segment range="10946-10963" color="#ffffff" type="standard"/><Segment name="-35" range="10964-10969" 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" consecutiveTranslationNumbering="1"><Segment range="10984-11005" 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="10" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4370-4386" 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="11" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="10891-10907" 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="12" name="lac operator" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="10915-10931" 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-Î²-D-thiogalactopyranoside (IPTG).&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>6fb109f6-5dd8-4110-9e12-e4042e89c4f3</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pKCcas9do</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Created UTC="9:59:44">2020.4.17</Created>
<LastModified UTC="11:38:14">2025.8.27</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y     K         
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   )<?xml version="1.0"?><CustomEnzymeSets/>
