LOCUS       Exported                9196 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 9196)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9196
                     /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..1347
                     /locus_tag="H3.2"
                     /label=H3.2
     misc_feature    1348..1392
                     /locus_tag="GS3Linker"
                     /label=GS3Linker
     misc_feature    1393..5541
                     /locus_tag="Cas9"
                     /label=Cas9
     misc_feature    5557..5610
                     /locus_tag="T2A"
                     /label=T2A
     misc_feature    5611..6210
                     /locus_tag="puroR"
                     /label=puroR
     rep_origin      6675..7130
                     /locus_tag="M13 origin"
                     /label=M13 origin
     rep_origin      6692..6998
                     /locus_tag="F1 ori"
                     /label=F1 ori
     rep_origin      8472..9154
                     /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 ccgcccgaac caagcagact
      961 gctcgtaagt ccacaggtgg gaaagccccc cgcaaacagc tggccacgaa agccgccagg
     1021 aaaagcgctc ccgctaccgg cggggtgaag aagcctcatc gctacaggcc cgggaccgtg
     1081 gcgcttcgag agattcgtcg ttatcagaag tcgaccgagc tgctcatccg gaagctgccc
     1141 ttccagaggt tggtgaggga gatcgcgcag gatttcaaaa ccgacctgag gtttcagagc
     1201 tcagccgtca tggcgctgca ggaggctagc gaagcgtacc tggtgggtct gttcgaagat
     1261 accaacctgt gtgccatcca cgctaagaga gtcaccatca tgcccaaaga catccagttg
     1321 gctcgccgga tacggggaga gagagctggt ggcggtggat ccggtggcgg tggaagtggt
     1381 ggcggtggat ctgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     1441 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     1501 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     1561 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     1621 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     1681 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     1741 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     1801 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     1861 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     1921 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     1981 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     2041 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     2101 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     2161 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     2221 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     2281 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     2341 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     2401 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     2461 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     2521 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     2581 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     2641 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     2701 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     2761 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     2821 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     2881 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     2941 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     3001 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     3061 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     3121 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     3181 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     3241 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     3301 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     3361 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     3421 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     3481 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     3541 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     3601 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     3661 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     3721 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     3781 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     3841 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     3901 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     3961 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     4021 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     4081 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     4141 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     4201 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     4261 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     4321 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     4381 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     4441 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     4501 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     4561 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     4621 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     4681 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     4741 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     4801 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     4861 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     4921 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     4981 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     5041 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     5101 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     5161 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     5221 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     5281 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     5341 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     5401 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     5461 acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc
     5521 ggccaggcaa aaaagaaaaa ggaattcggc agtggagagg gcagaggaag tctgctaaca
     5581 tgcggtgacg tcgaggagaa tcctggccca atgaccgagt acaagcccac ggtgcgcctc
     5641 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5701 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5761 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5821 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5881 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5941 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc
     6001 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     6061 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     6121 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     6181 tggtgcatga cccgcaagcc cggtgcctga gaattctaac tagagctcgc tgatcagcct
     6241 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6301 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6361 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6421 attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta gtgatggagt
     6481 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     6541 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     6601 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     6661 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     6901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     7021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7081 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     7141 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     7201 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     7261 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     7321 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     7381 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7741 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8041 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     8101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     8161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8221 cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     8701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     9001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     9061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     9121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     9181 gccttttgct cacatg
//
