LOCUS       plasmid                 8504 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     CDS             complement(698..3196)
                     /note="pLannotate"
                     /label="Cas9(C)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             complement(3197..4798)
                     /note="pLannotate"
                     /label="Cas9(N)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             complement(6987..7847)
                     /note="pLannotate"
                     /label="AmpR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     enhancer        complement(5440..5725)
                     /note="pLannotate"
                     /label="CMV enhancer"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
     intron          complement(4936..5163)
                     /note="pLannotate"
                     /label="hybrid intron"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="intron"
     repeat_region   complement(260..400)
                     /note="pLannotate"
                     /label="AAV2 ITR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="repeat_region"
     polyA_signal    complement(505..616)
                     /note="pLannotate"
                     /label="bGH poly(A) signal"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="polyA_signal"
     promoter        complement(7848..7952)
                     /note="pLannotate"
                     /label="AmpR promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        complement(5822..5897)
                     /note="pLannotate"
                     /label="gRNA scaffold"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="misc_RNA"
     rep_origin      complement(6228..6816)
                     /note="pLannotate"
                     /label="ori"
                     /database="snapgene"
                     /identity="99.8"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     rep_origin      complement(join(8234..8504,1..185))
                     /note="pLannotate"
                     /label="f1 ori"
                     /database="snapgene"
                     /identity="98.9"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     promoter        complement(5164..5438)
                     /note="pLannotate"
                     /label="chicken β-actin promoter"
                     /database="snapgene"
                     /identity="98.9"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     promoter        complement(5926..6166)
                     /note="pLannotate"
                     /label="U6 promoter"
                     /database="snapgene"
                     /identity="99.6"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        complement(5772..5815)
                     /note="pLannotate"
                     /label="gRNA scaffold (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="57.9"
                     /fragment="True"
                     /other="misc_RNA"
     misc_feature    204..255
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="36.9"
                     /fragment="True"
                     /other="misc_feature"
     misc_feature    8184..8233
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="35.5"
                     /fragment="True"
                     /other="misc_feature"
     CDS             8047..8082
                     /note="pLannotate"
                     /label="rop (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="18.8"
                     /fragment="True"
                     /other="CDS"
ORIGIN
        1 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
       61 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
      121 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      181 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      241 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
      301 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
      361 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc cccagcatgc
      421 ctgctattct cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
      481 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
      541 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
      601 caactagaag gcacagtcga ggctgatcag cgagctctag gaattcttac tttttctttt
      661 ttgcctggcc ggcctttttc gtggccgccg gccttttgtc gcctcccagc tgagacaggt
      721 cgatccgtgt ctcgtacagg ccggtgatgc tctggtggat cagggtggcg tccagcacct
      781 ctttggtgct ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg aaggcggcag
      841 gggctcccag attggtcagg gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg
      901 gcttatcccg gtgcttgttg taggcggaca gcactttgtc cagattagcg tcggccagga
      961 tcactctctt ggagaactcg ctgatctgct cgatgatctc gtccaggtag tgcttgtgct
     1021 gttccacaaa cagctgtttc tgctcattat cctcggggga gcccttcagc ttctcatagt
     1081 ggctggccag gtacaggaag ttcacatatt tggagggcag ggccagttcg tttcccttct
     1141 gcagttcgcc ggcagaggcc agcattctct tccggccgtt ttccagctcg aacagggagt
     1201 acttaggcag cttgatgatc aggtcctttt tcacttcttt gtagcccttg gcttccagaa
     1261 agtcgatggg attcttctcg aagctgcttc tttccatgat ggtgatcccc agcagctctt
     1321 tcacactctt cagtttcttg gacttgccct tttccacttt ggccaccacc agcacagaat
     1381 aggccacggt ggggctgtcg aagccgccgt acttcttagg gtcccagtcc ttctttctgg
     1441 cgatcagctt atcgctgttc ctcttgggca ggatagactc tttgctgaag ccgcctgtct
     1501 gcacctcggt ctttttcacg atattcactt ggggcatgct cagcactttc cgcacggtgg
     1561 caaaatcccg gcccttatcc cacacgatct ccccggtttc gccgtttgtc tcgatcagag
     1621 gccgcttccg gatctcgccg ttggccaggg taatctcggt cttgaaaaag ttcatgatgt
     1681 tgctgtagaa gaagtacttg gcggtagcct tgccgatttc ctgctcgctc ttggcgatca
     1741 tcttccgcac gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt
     1801 actttttgat cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt
     1861 agttgttgat ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct
     1921 tggacttcag ggtgatcact ttcacttccc ggatcagctt gtcattctcg tcgtacttag
     1981 tgttcatccg ggagtccagg atctgtgcca cgtgctttgt gatctgccgg gtttccacca
     2041 gctgtctctt gatgaagccg gccttatcca gttcgctcag gccgcctctc tcggccttgg
     2101 tcagattgtc gaactttctc tgggtaatca gcttggcgtt cagcagctgc cgccagtagt
     2161 tcttcatctt cttcacgacc tcttcggagg gcacgttgtc gctcttgccc cggttcttgt
     2221 cgcttctggt cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga
     2281 tatggtccac atcgtagtcg gacagccggt tgatgtccag ttcctggtcc acgtacatat
     2341 cccgcccatt ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca
     2401 cggggtgttc tttcaggatc tggctgccca gctctttgat gccctcttcg atccgcttca
     2461 ttctctcgcg gctgttcttc tgtcccttct gggtggtctg gttctctctg gccatttcga
     2521 tcacgatgtt ctcgggcttg tgccggccca tcactttcac gagctcgtcc accaccttca
     2581 ctgtctgcag gatgcccttc ttaatggcgg ggctgccggc cagattggca atgtgctcgt
     2641 gcaggctatc gccctggccg gacacctggg ctttctggat gtcctcttta aaggtcaggc
     2701 tgtcgtcgtg gatcagctgc atgaagtttc tgttggcgaa gccgtcggac ttcaggaaat
     2761 ccaggattgt cttgccggac tgcttgtccc ggatgccgtt gatcagcttc cggctcagcc
     2821 tgccccagcc ggtgtatctc cgccgcttca gctgcttcat cactttgtcg tcgaacaggt
     2881 gggcataggt tttcagccgt tcctcgatca tctctctgtc ctcaaacagt gtcagggtca
     2941 gcacgatatc ttccagaatg tcctcgtttt cctcattgtc caggaagtcc ttgtccttga
     3001 taattttcag cagatcgtgg tatgtgccca gggaggcgtt gaaccgatct tccacgccgg
     3061 agatttccac ggagtcgaag cactcgattt tcttgaagta gtcctctttc agctgcttca
     3121 cggtcacttt ccggttggtc ttgaacagca ggtccacgat ggcctttttc tgctcgccgc
     3181 tcaggaaggc gggctttctc attccctcgg tcacgtattt cactttggtc agctcgttat
     3241 acacggtgaa gtactcgtac agcaggctgt gcttgggcag caccttctcg ttgggcaggt
     3301 tcttatcgaa gttggtcatc cgctcgatga agctctgggc ggaagcgccc ttgtccacca
     3361 cttcctcgaa gttccagggg gtgatggttt cctcgctctt tctggtcatc caggcgaatc
     3421 tgctgtttcc cctggccaga gggcccacgt agtaggggat gcggaaggtc aggatcttct
     3481 cgatcttttc ccggttgtcc ttcaggaatg ggtaaaaatc ttcctgccgc cgcagaatgg
     3541 cgtgcagctc tcccaggtgg atctggtggg ggatgctgcc gttgtcgaag gtccgctgct
     3601 tccgcagcag gtcctctctg ttcagcttca cgagcagttc ctcggtgccg tccatctttt
     3661 ccaggatggg cttgatgaac ttgtagaact cttcctggct ggctccgccg tcaatgtagc
     3721 cggcgtagcc gttcttgctc tggtcgaaga aaatctcttt gtacttctca ggcagctgct
     3781 gccgcacgag agctttcagc agggtcaggt cctggtggtg ctcgtcgtat ctcttgatca
     3841 tagaggcgct caggggggcc ttggtgatct cggtgttcac tctcaggatg tcgctcagca
     3901 ggatggcgtc ggacaggttc ttggcggcca gaaacaggtc ggcgtactgg tcgccgatct
     3961 gggccagcag gttgtccagg tcgtcgtcgt aggtgtcctt gctcagctgc agtttggcat
     4021 cctcggccag gtcgaagttg ctcttgaagt tgggggtcag gcccaggctc agggcaatca
     4081 ggtttccgaa caggccattc ttcttctcgc cgggcagctg ggcgatcaga ttttccagcc
     4141 gtctgctctt gctcagtctg gcagacagga tggccttggc gtccacgccg ctggcgttga
     4201 tggggttttc ctcgaacagc tggttgtagg tctgcaccag ctggatgaac agcttgtcca
     4261 cgtcgctgtt gtcggggttc aggtcgccct cgatcaggaa gtggccccgg aacttgatca
     4321 tgtgggccag ggccagatag atcagccgca ggtcggcctt gtcggtgctg tccaccagtt
     4381 tctttctcag gtggtagatg gtggggtact tctcgtggta ggccacctcg tccacgatgt
     4441 tgccgaagat ggggtgccgc tcgtgcttct tatcctcttc caccaggaag gactcttcca
     4501 gtctgtggaa gaagctgtcg tccaccttgg ccatctcgtt gctgaagatc tcttgcagat
     4561 agcagatccg gttcttccgt ctggtgtatc ttcttctggc ggttctcttc agccgggtgg
     4621 cctcggctgt ttcgccgctg tcgaacagca gggctccgat caggttcttc ttgatgctgt
     4681 gccggtcggt gttgcccagc accttgaatt tcttgctggg caccttgtac tcgtcggtga
     4741 tcacggccca gcccacagag ttggtgccga tgtccaggcc gatgctgtac ttcttgtcgg
     4801 ctgctgggac tccgtggata ccgaccttcc gcttcttctt tggggccatc ttatcgtcat
     4861 cgtctttgta atcaatatca tgatccttgt agtctccgtc gtggtcctta tagtccatgg
     4921 tggcaccggt ccaacctgaa aaaaagtgat ttcaggcagg tgctccaggt aattaaacat
     4981 taatacccca ccaaccaacc atcccttaaa cccttacctc ttgctcagct aattacagcc
     5041 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
     5101 gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac
     5161 tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa
     5221 aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc
     5281 gccccgcccc tcgccccgcc ccgccccgcc tggcgcgcgc cccccccccc cccgccccca
     5341 tcgctgcaca aaataattaa aaaataaata aatacaaaat tgggggtggg gaggggggga
     5401 gatggggaga gtgaagcaga acgtggggct cacctcgacc atggtaatag cgatgactaa
     5461 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cacaatgcca
     5521 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     5581 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     5641 ccctattggc gttactattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5701 gccatttacc gtaagttatg taacgggtac ctctagagcc atttgtctgc agaattggcg
     5761 cacgcgctaa aaacggacta gccttatttt aacttgctat ttctagctct aaaacaaaaa
     5821 agcaccgact cggtgccact ttttcaagtt gataacggac tagccttatt ttaacttgct
     5881 atttctagct ctaaaacatc ctgtccctag tggccccggt gtttcgtcct ttccacaaga
     5941 tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt
     6001 taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat
     6061 tttgtactaa tatctttgtg tttacagtca aattaattcc aattatctct ctaacagcct
     6121 tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcacat gtgagcaaaa
     6181 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6241 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6301 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6361 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6421 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6481 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6541 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6601 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6661 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6721 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6781 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6841 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6901 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6961 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7021 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7081 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggct tccacgctca
     7141 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7201 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7261 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7321 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7381 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7441 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7501 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7561 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7621 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7681 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7741 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7801 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7861 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7921 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7981 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8041 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     8101 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     8161 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     8221 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     8281 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
     8341 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     8401 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     8461 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaa
//
