LOCUS       Exported                9217 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 9217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9217
                     /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..1368
                     /locus_tag="H2A.X"
                     /label=H2A.X
     misc_feature    1339..1340
                     /locus_tag="K134L"
                     /label=K134L
     misc_feature    1369..1413
                     /locus_tag="GS3Linker"
                     /label=GS3Linker
     misc_feature    1414..5562
                     /locus_tag="Cas9"
                     /label=Cas9
     misc_feature    5578..5631
                     /locus_tag="T2A"
                     /label=T2A
     misc_feature    5632..6231
                     /locus_tag="puroR"
                     /label=puroR
     rep_origin      6696..7151
                     /locus_tag="M13 origin"
                     /label=M13 origin
     rep_origin      6713..7019
                     /locus_tag="F1 ori"
                     /label=F1 ori
     rep_origin      8493..9175
                     /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 cctcgggccg cggcaagact
      961 ggcggcaagg cccgcgccaa ggccaagtcg cgctcgtcgc gcgccggcct ccagttccca
     1021 gtgggccgtg tacaccggct gctgcggaag ggccactacg ccgagcgcgt tggcgccggc
     1081 gcgccagtgt acctggcggc agtgctggag tacctcaccg ctgagatcct ggagctggcg
     1141 ggcaatgcgg cccgcgacaa caagaagacg cgaatcatcc cccgccacct gcagctggcc
     1201 atccgcaacg acgaggagct caacaagctg ctgggcggcg tgacgatcgc ccagggaggc
     1261 gtcctgccca acatccaggc cgtgctgctg cccaagaaga ccagcgccac cgtggggccg
     1321 aaggcgccct cgggcggctt gaaggccacc caggcctccc aggagtacgg tggcggtgga
     1381 tccggtggcg gtggaagtgg tggcggtgga tctgacaaga agtacagcat cggcctggac
     1441 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     1501 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     1561 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     1621 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     1681 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     1741 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     1801 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     1861 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     1921 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     1981 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     2041 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     2101 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     2161 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     2221 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     2281 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     2341 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     2401 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     2461 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     2521 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     2581 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     2641 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     2701 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     2761 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     2821 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     2881 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     2941 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     3001 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     3061 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     3121 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     3181 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     3241 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     3301 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     3361 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     3421 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     3481 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     3541 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     3601 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     3661 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     3721 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     3781 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     3841 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     3901 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     3961 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     4021 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     4081 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     4141 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     4201 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     4261 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     4321 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     4381 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     4441 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     4501 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     4561 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     4621 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     4681 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     4741 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     4801 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     4861 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     4921 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     4981 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     5041 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     5101 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     5161 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     5221 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     5281 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     5341 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     5401 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     5461 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg
     5521 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa aggaattcgg cagtggagag
     5581 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatgaccgag
     5641 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     5701 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     5761 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5821 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5881 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5941 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6001 ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6061 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6121 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6181 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agaattctaa
     6241 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     6301 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     6361 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     6421 ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc
     6481 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     6541 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     6601 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     6661 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     6721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6901 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     7021 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     7081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     7141 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7201 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     7261 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     7321 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     7381 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     7441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     7501 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     7561 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     7621 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7681 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7741 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7801 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7861 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7921 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7981 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     8041 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     8101 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     8161 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     8221 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     8281 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     8341 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     8401 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     8461 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     8521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     8581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     8641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8701 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8941 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     9001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     9061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     9121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     9181 acggttcctg gccttttgct ggccttttgc tcacatg
//
