LOCUS       plasmid                 8506 bp    DNA     circular SYN 03-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     CDS             5363..7861
                     /note="pLannotate"
                     /label="Cas9(C)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             3761..5362
                     /note="pLannotate"
                     /label="Cas9(N)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             710..1570
                     /note="pLannotate"
                     /label="AmpR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     enhancer        2833..3118
                     /note="pLannotate"
                     /label="CMV enhancer"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
     intron          3396..3623
                     /note="pLannotate"
                     /label="hybrid intron"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="intron"
     repeat_region   8159..8299
                     /note="pLannotate"
                     /label="AAV2 ITR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="repeat_region"
     polyA_signal    7943..8054
                     /note="pLannotate"
                     /label="bGH poly(A) signal"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="polyA_signal"
     promoter        605..709
                     /note="pLannotate"
                     /label="AmpR promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        2661..2736
                     /note="pLannotate"
                     /label="gRNA scaffold"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="misc_RNA"
     rep_origin      1741..2329
                     /note="pLannotate"
                     /label="ori"
                     /database="snapgene"
                     /identity="99.8"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     rep_origin      join(8374..8506,1..323)
                     /note="pLannotate"
                     /label="f1 ori"
                     /database="snapgene"
                     /identity="98.9"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     promoter        3120..3395
                     /note="pLannotate"
                     /label="chicken β-actin promoter"
                     /database="snapgene"
                     /identity="99.3"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     promoter        2391..2631
                     /note="pLannotate"
                     /label="U6 promoter"
                     /database="snapgene"
                     /identity="99.6"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        2743..2786
                     /note="pLannotate"
                     /label="gRNA scaffold (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="57.9"
                     /fragment="True"
                     /other="misc_RNA"
     misc_feature    complement(8304..8355)
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="36.9"
                     /fragment="True"
                     /other="misc_feature"
     misc_feature    complement(324..373)
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="35.5"
                     /fragment="True"
                     /other="misc_feature"
     CDS             complement(475..510)
                     /note="pLannotate"
                     /label="rop (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="18.8"
                     /fragment="True"
                     /other="CDS"
ORIGIN
        1 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
       61 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
      121 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
      181 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
      241 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
      301 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
      361 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      421 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      481 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
      541 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      601 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      661 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      721 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      781 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      841 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      901 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      961 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1021 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1081 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1141 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1201 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1261 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1321 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1381 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     1441 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1501 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1561 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1621 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1801 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
     2401 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     2461 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     2521 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     2581 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     2641 ttccaggtct gtggggcagt gttttagagc tagaaatagc aagttaaaat aaggctagtc
     2701 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga gctagaaata
     2761 gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca gacaaatggc
     2821 tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2881 cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac gtcaatgggt
     2941 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3001 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc agtacatgac
     3061 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     3121 cgaggtgagc cccacgttct gcttcactct ccccatctcc ccccctcccc acccccaatt
     3181 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggc
     3241 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     3301 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     3361 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     3421 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     3481 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga
     3541 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc
     3601 tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata aggaccacga
     3661 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa
     3721 gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg
     3781 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     3841 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     3901 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     3961 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     4021 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     4081 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     4141 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     4201 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     4261 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     4321 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     4381 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     4441 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     4501 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     4561 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     4621 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     4681 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     4741 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     4801 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     4861 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     4921 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     4981 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     5041 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     5101 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     5161 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     5221 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     5281 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     5341 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     5401 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     5461 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     5521 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     5581 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     5641 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     5701 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     5761 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     5821 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     5881 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     5941 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     6001 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     6061 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     6121 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     6181 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     6241 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     6301 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     6361 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     6421 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     6481 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     6541 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     6601 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     6661 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     6721 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     6781 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     6841 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     6901 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     6961 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     7021 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     7081 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     7141 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     7201 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     7261 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     7321 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     7381 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     7441 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     7501 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     7561 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     7621 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     7681 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     7741 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
     7801 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     7861 caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagt aagaattcct
     7921 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     7981 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     8041 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     8101 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag
     8161 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     8221 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     8281 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     8341 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     8401 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     8461 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctt
//
