LOCUS       plasmid                 8505 bp    DNA     circular SYN 09-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     CDS             5541..8039
                     /note="pLannotate"
                     /label="Cas9(C)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             3939..5540
                     /note="pLannotate"
                     /label="Cas9(N)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             889..1749
                     /note="pLannotate"
                     /label="AmpR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     enhancer        3012..3297
                     /note="pLannotate"
                     /label="CMV enhancer"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
     intron          3574..3801
                     /note="pLannotate"
                     /label="hybrid intron"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="intron"
     repeat_region   8337..8477
                     /note="pLannotate"
                     /label="AAV2 ITR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="repeat_region"
     polyA_signal    8121..8232
                     /note="pLannotate"
                     /label="bGH poly(A) signal"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="polyA_signal"
     promoter        784..888
                     /note="pLannotate"
                     /label="AmpR promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        2840..2915
                     /note="pLannotate"
                     /label="gRNA scaffold"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="misc_RNA"
     rep_origin      1920..2508
                     /note="pLannotate"
                     /label="ori"
                     /database="snapgene"
                     /identity="99.8"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     rep_origin      47..502
                     /note="pLannotate"
                     /label="f1 ori"
                     /database="snapgene"
                     /identity="98.9"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     promoter        3299..3573
                     /note="pLannotate"
                     /label="chicken β-actin promoter"
                     /database="snapgene"
                     /identity="98.9"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     promoter        2570..2810
                     /note="pLannotate"
                     /label="U6 promoter"
                     /database="snapgene"
                     /identity="99.6"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        2922..2965
                     /note="pLannotate"
                     /label="gRNA scaffold (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="57.9"
                     /fragment="True"
                     /other="misc_RNA"
     misc_feature    complement(join(8482..8505,1..28))
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="36.9"
                     /fragment="True"
                     /other="misc_feature"
     misc_feature    complement(503..552)
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="35.5"
                     /fragment="True"
                     /other="misc_feature"
     CDS             complement(654..689)
                     /note="pLannotate"
                     /label="rop (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="18.8"
                     /fragment="True"
                     /other="CDS"
ORIGIN
        1 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
       61 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      121 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      181 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      241 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
      301 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      361 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
      421 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      481 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
      541 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      601 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      661 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
      721 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1561 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     1621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
     2581 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     2641 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     2701 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     2761 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt
     2821 ggggtgtcgg aattcaaaag ttttagagct agaaatagca agttaaaata aggctagtcc
     2881 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag
     2941 caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct
     3001 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3061 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
     3121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     3181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
     3241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     3301 gaggtgagcc ccacgttctg cttcactctc cccatctccc cccctcccca cccccaattt
     3361 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
     3421 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     3481 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     3541 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
     3601 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     3661 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
     3721 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
     3781 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg
     3841 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
     3901 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
     3961 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     4021 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     4081 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     4141 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     4201 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     4261 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     4321 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     4381 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     4441 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     4501 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     4561 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     4621 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     4681 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     4741 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     4801 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     4861 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     4921 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     4981 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     5041 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     5101 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     5161 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     5221 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     5281 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     5341 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     5401 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     5461 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     5521 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     5581 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     5641 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     5701 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     5761 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     5821 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     5881 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     5941 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     6001 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     6061 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     6121 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     6181 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     6241 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     6301 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     6361 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     6421 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     6481 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     6541 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     6601 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     6661 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     6721 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     6781 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     6841 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     6901 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     6961 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     7021 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     7081 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     7141 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     7201 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     7261 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     7321 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     7381 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     7441 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     7501 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     7561 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     7621 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     7681 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     7741 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     7801 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     7861 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     7921 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     7981 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     8041 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag
     8101 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     8161 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     8221 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     8281 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga
     8341 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     8401 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     8461 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttac
//
