LOCUS       plasmid                 8506 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     CDS             join(7137..8506,1..1129)
                     /note="pLannotate"
                     /label="Cas9(C)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             5535..7136
                     /note="pLannotate"
                     /label="Cas9(N)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     CDS             2484..3344
                     /note="pLannotate"
                     /label="AmpR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="CDS"
     enhancer        4607..4892
                     /note="pLannotate"
                     /label="CMV enhancer"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="enhancer"
     intron          5170..5397
                     /note="pLannotate"
                     /label="hybrid intron"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="intron"
     repeat_region   1427..1567
                     /note="pLannotate"
                     /label="AAV2 ITR"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="repeat_region"
     polyA_signal    1211..1322
                     /note="pLannotate"
                     /label="bGH poly(A) signal"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="polyA_signal"
     promoter        2379..2483
                     /note="pLannotate"
                     /label="AmpR promoter"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        4435..4510
                     /note="pLannotate"
                     /label="gRNA scaffold"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="100.0"
                     /fragment="False"
                     /other="misc_RNA"
     rep_origin      3515..4103
                     /note="pLannotate"
                     /label="ori"
                     /database="snapgene"
                     /identity="99.8"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     rep_origin      1642..2097
                     /note="pLannotate"
                     /label="f1 ori"
                     /database="snapgene"
                     /identity="98.9"
                     /match_length="100.0"
                     /fragment="False"
                     /other="rep_origin"
     promoter        4894..5169
                     /note="pLannotate"
                     /label="chicken β-actin promoter"
                     /database="snapgene"
                     /identity="99.3"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     promoter        4165..4405
                     /note="pLannotate"
                     /label="U6 promoter"
                     /database="snapgene"
                     /identity="99.6"
                     /match_length="100.0"
                     /fragment="False"
                     /other="promoter"
     misc_RNA        4517..4560
                     /note="pLannotate"
                     /label="gRNA scaffold (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="57.9"
                     /fragment="True"
                     /other="misc_RNA"
     misc_feature    complement(1572..1623)
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="36.9"
                     /fragment="True"
                     /other="misc_feature"
     misc_feature    complement(2098..2147)
                     /note="pLannotate"
                     /label="bom (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="35.5"
                     /fragment="True"
                     /other="misc_feature"
     CDS             complement(2249..2284)
                     /note="pLannotate"
                     /label="rop (fragment)"
                     /database="snapgene"
                     /identity="100.0"
                     /match_length="18.8"
                     /fragment="True"
                     /other="CDS"
ORIGIN
        1 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
       61 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
      121 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
      181 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
      241 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
      301 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
      361 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
      421 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
      481 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
      541 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
      601 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
      661 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
      721 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
      781 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
      841 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
      901 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
      961 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     1021 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     1081 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc
     1141 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag agctcgctga
     1201 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1261 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1321 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1381 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg
     1441 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     1501 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     1561 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     1621 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1681 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     1741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     1861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     1981 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     2041 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     2101 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2161 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2221 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2281 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2341 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2401 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2461 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2521 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2581 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2641 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2701 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2761 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2821 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2881 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2941 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3001 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3061 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3181 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     3241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4141 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
     4201 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     4261 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     4321 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     4381 gctttatata tcttgtggaa aggacgaaac accgtagcta cacagtaagt agctgtttta
     4441 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     4501 gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     4561 tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt acataactta
     4621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
     4681 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4741 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4801 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
     4861 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     4921 ctctccccat ctccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
     4981 ttgtgcagcg atgggggcgg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     5041 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     5101 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     5161 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     5221 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     5281 ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt
     5341 ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt
     5401 ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat catgatattg
     5461 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
     5521 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     5581 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     5641 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     5701 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     5761 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     5821 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     5881 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     5941 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     6001 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     6061 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     6121 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     6181 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     6241 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     6301 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     6361 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     6421 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     6481 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     6541 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     6601 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     6661 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     6721 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     6781 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     6841 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     6901 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     6961 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     7021 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     7081 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     7141 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     7201 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     7261 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     7321 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     7381 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     7441 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     7501 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     7561 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     7621 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     7681 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     7741 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     7801 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     7861 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     7921 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     7981 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     8041 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     8101 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     8161 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     8221 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     8281 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     8341 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     8401 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     8461 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgt
//
