LOCUS       Exported                9133 bp ds-DNA     circular SYN 03-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9133)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9133
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    12..810
                     /locus_tag="CBh"
                     /label=CBh
     misc_feature    766..785
                     /locus_tag="CBhFW1"
                     /label=CBhFW1
     misc_feature    823..900
                     /locus_tag="3xFLAG"
                     /label=3xFLAG
     misc_feature    901..915
                     /locus_tag="NLS"
                     /label=NLS
     misc_feature    943..1284
                     /locus_tag="H2A.B"
                     /label=H2A.B
     misc_feature    1285..1329
                     /locus_tag="GS3Linker"
                     /label=GS3Linker
     misc_feature    1330..5478
                     /locus_tag="Cas9"
                     /label=Cas9
     misc_feature    5494..5547
                     /locus_tag="T2A"
                     /label=T2A
     misc_feature    5548..6147
                     /locus_tag="puroR"
                     /label=puroR
     rep_origin      6612..7067
                     /locus_tag="M13 origin"
                     /label=M13 origin
     rep_origin      6629..6935
                     /locus_tag="F1 ori"
                     /label=F1 ori
     rep_origin      8409..9091
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
ORIGIN
        1 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
       61 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
      121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
      241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
      301 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
      361 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
      421 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      481 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      541 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgac gctgccttcg
      601 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      661 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa
      721 gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca
      781 cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggacta taaggaccac
      841 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca
      901 aagaagaagc ggaaggtcgg gatccacgga gtccctgcag ccccgaggag gaggagacgc
      961 cgagggtcct ccggtgctgg cggccggggg cggacctgct ctcgcaccgt ccgagcggag
     1021 ctttcgtttt cagtgagcca ggtggagcgc agtctacggg agggccacta cgctcagcgc
     1081 ctgagtcgca cggcgccggt ctacctcgct gcggttattg agtacctgac ggccaaggtc
     1141 ccggagctgg cgggcaacga ggcccagaac agcggagagc ggaacatcac tcccctgctg
     1201 ctggacatgg tggttcacaa cgacaggcta ctgagcaccc ttttcaacac gaccaccatc
     1261 tctcaagtgg cccctggcga ggacggtggc ggtggatccg gtggcggtgg aagtggtggc
     1321 ggtggatctg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     1381 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     1441 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     1501 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     1561 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     1621 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     1681 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     1741 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     1801 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     1861 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     1921 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     1981 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     2041 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     2101 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     2161 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     2221 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     2281 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     2341 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     2401 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     2461 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     2521 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     2581 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     2641 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     2701 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     2761 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     2821 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     2881 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     2941 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     3001 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     3061 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     3121 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     3181 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     3241 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     3301 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     3361 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     3421 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     3481 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     3541 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     3601 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     3661 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     3721 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     3781 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     3841 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     3901 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     3961 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     4021 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     4081 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     4141 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     4201 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     4261 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     4321 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     4381 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     4441 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     4501 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     4561 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     4621 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     4681 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     4741 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     4801 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     4861 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     4921 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     4981 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     5041 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     5101 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     5161 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     5221 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     5281 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     5341 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     5401 cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc
     5461 caggcaaaaa agaaaaagga attcggcagt ggagagggca gaggaagtct gctaacatgc
     5521 ggtgacgtcg aggagaatcc tggcccaatg accgagtaca agcccacggt gcgcctcgcc
     5581 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5641 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5701 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5761 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     5821 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     5881 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg
     5941 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6001 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6061 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6121 tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag agctcgctga tcagcctcga
     6181 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6241 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6301 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6361 gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg atggagttgg
     6421 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     6481 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     6541 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     6601 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     6841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     6961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7021 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     7081 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     7141 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     7201 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     7261 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     7321 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7381 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7441 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7501 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7561 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7621 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7681 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7741 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7801 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7861 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7921 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7981 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8041 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8101 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8161 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8221 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8281 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8341 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8401 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8461 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8521 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8581 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     8641 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8701 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8761 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8821 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8881 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8941 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9001 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9061 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9121 ttttgctcac atg
//
