LOCUS       .                       8220 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     CDS             407..430
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             437..457
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             482..4402
                     /label="AsCpf1"
                     /note="CRISPR-associated protein from Acidaminococcus sp.
                     BV3L6 (Zetsche et al., 2015)"
     CDS             4520..5233
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     polyA_signal    5267..5474
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5483..5623
                     /label="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
     rep_origin      5698..6153
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6435..6539
                     /label="AmpR promoter"
     CDS             6540..7397
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      7571..8159
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cactctggcc tctgattggt ccaaggaagg ctggggggca ggacgggagg cgaaaaccct
       61 ggaatattcc cgacctggca gcctcatcga gctcggtgat tggctcagaa gggaaaaggc
      121 gggtctccgt gacgacttat aaaagcccag gggcaagcgg tccggataac ggctagcctg
      181 aggagctgct gcgacagtcc actacctttt tcgagagtga ctcccgttgt cccaaggctt
      241 cccagagcga acctgtgcgg ctgcaggcac cggcgcgtcg agtttccggc gtccggaagg
      301 accgagctct tctcgcggat ccagtgttcc gtttccagag aaacaccgga ccggtgccac
      361 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
      421 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
      481 catgacacag ttcgagggct ttaccaacct gtatcaggtg agcaagacac tgcggtttga
      541 gctgatccca cagggcaaga ccctgaagca catccaggag cagggcttca tcgaggagga
      601 caaggcccgc aatgatcact acaaggagct gaagcccatc atcgatcgga tctacaagac
      661 ctatgccgac cagtgcctgc agctggtgca gctggattgg gagaacctga gcgccgccat
      721 cgactcctat agaaaggaga aaaccgagga gacaaggaac gccctgatcg aggagcaggc
      781 cacatatcgc aatgccatcc acgactactt catcggccgg acagacaacc tgaccgatgc
      841 catcaataag agacacgccg agatctacaa gggcctgttc aaggccgagc tgtttaatgg
      901 caaggtgctg aagcagctgg gcaccgtgac cacaaccgag cacgagaacg ccctgctgcg
      961 gagcttcgac aagtttacaa cctacttctc cggcttttat gagaacagga agaacgtgtt
     1021 cagcgccgag gatatcagca cagccatccc acaccgcatc gtgcaggaca acttccccaa
     1081 gtttaaggag aattgtcaca tcttcacacg cctgatcacc gccgtgccca gcctgcggga
     1141 gcactttgag aacgtgaaga aggccatcgg catcttcgtg agcacctcca tcgaggaggt
     1201 gttttccttc cctttttata accagctgct gacacagacc cagatcgacc tgtataacca
     1261 gctgctggga ggaatctctc gggaggcagg caccgagaag atcaagggcc tgaacgaggt
     1321 gctgaatctg gccatccaga agaatgatga gacagcccac atcatcgcct ccctgccaca
     1381 cagattcatc cccctgttta agcagatcct gtccgatagg aacaccctgt ctttcatcct
     1441 ggaggagttt aagagcgacg aggaagtgat ccagtccttc tgcaagtaca agacactgct
     1501 gagaaacgag aacgtgctgg agacagccga ggccctgttt aacgagctga acagcatcga
     1561 cctgacacac atcttcatca gccacaagaa gctggagaca atcagcagcg ccctgtgcga
     1621 ccactgggat acactgagga atgccctgta tgagcggaga atctccgagc tgacaggcaa
     1681 gatcaccaag tctgccaagg agaaggtgca gcgcagcctg aagcacgagg atatcaacct
     1741 gcaggagatc atctctgccg caggcaagga gctgagcgag gccttcaagc agaaaaccag
     1801 cgagatcctg tcccacgcac acgccgccct ggatcagcca ctgcctacaa ccctgaagaa
     1861 gcaggaggag aaggagatcc tgaagtctca gctggacagc ctgctgggcc tgtaccacct
     1921 gctggactgg tttgccgtgg atgagtccaa cgaggtggac cccgagttct ctgcccggct
     1981 gaccggcatc aagctggaga tggagccttc tctgagcttc tacaacaagg ccagaaatta
     2041 tgccaccaag aagccctact ccgtggagaa gttcaagctg aactttcaga tgcctacact
     2101 ggcctctggc tgggacgtga ataaggagaa gaacaatggc gccatcctgt ttgtgaagaa
     2161 cggcctgtac tatctgggca tcatgccaaa gcagaagggc aggtataagg ccctgagctt
     2221 cgagcccaca gagaaaacca gcgagggctt tgataagatg tactatgact acttccctga
     2281 tgccgccaag atgatcccaa agtgcagcac ccagctgaag gccgtgacag cccactttca
     2341 gacccacaca acccccatcc tgctgtccaa caatttcatc gagcctctgg agatcacaaa
     2401 ggagatctac gacctgaaca atcctgagaa ggagccaaag aagtttcaga cagcctacgc
     2461 caagaaaacc ggcgaccaga agggctacag agaggccctg tgcaagtgga tcgacttcac
     2521 aagggatttt ctgtccaagt ataccaagac aacctctatc gatctgtcta gcctgcggcc
     2581 atcctctcag tataaggacc tgggcgagta ctatgccgag ctgaatcccc tgctgtacca
     2641 catcagcttc cagagaatcg ccgagaagga gatcatggat gccgtggaga caggcaagct
     2701 gtacctgttc cagatctata acaaggactt tgccaagggc caccacggca agcctaatct
     2761 gcacacactg tattggaccg gcctgttttc tccagagaac ctggccaaga caagcatcaa
     2821 gctgaatggc caggccgagc tgttctaccg ccctaagtcc aggatgaaga ggatggcaca
     2881 ccggctggga gagaagatgc tgaacaagaa gctgaaggat cagaaaaccc caatccccga
     2941 caccctgtac caggagctgt acgactatgt gaatcacaga ctgtcccacg acctgtctga
     3001 tgaggccagg gccctgctgc ccaacgtgat caccaaggag gtgtctcacg agatcatcaa
     3061 ggataggcgc tttaccagcg acaagttctt tttccacgtg cctatcacac tgaactatca
     3121 ggccgccaat tccccatcta agttcaacca gagggtgaat gcctacctga aggagcaccc
     3181 cgagacacct atcatcggca tcgatcgggg cgagagaaac ctgatctata tcacagtgat
     3241 cgactccacc ggcaagatcc tggagcagcg gagcctgaac accatccagc agtttgatta
     3301 ccagaagaag ctggacaaca gggagaagga gagggtggca gcaaggcagg cctggtctgt
     3361 ggtgggcaca atcaaggatc tgaagcaggg ctatctgagc caggtcatcc acgagatcgt
     3421 ggacctgatg atccactacc aggccgtggt ggtgctggag aacctgaatt tcggctttaa
     3481 gagcaagagg accggcatcg ccgagaaggc cgtgtaccag cagttcgaga agatgctgat
     3541 cgataagctg aattgcctgg tgctgaagga ctatccagca gagaaagtgg gaggcgtgct
     3601 gaacccatac cagctgacag accagttcac ctcctttgcc aagatgggca cccagtctgg
     3661 cttcctgttt tacgtgcctg ccccatatac atctaagatc gatcccctga ccggcttcgt
     3721 ggaccccttc gtgtggaaaa ccatcaagaa tcacgagagc cgcaagcact tcctggaggg
     3781 cttcgacttt ctgcactacg acgtgaaaac cggcgacttc atcctgcact ttaagatgaa
     3841 cagaaatctg tccttccaga ggggcctgcc cggctttatg cctgcatggg atatcgtgtt
     3901 cgagaagaac gagacacagt ttgacgccaa gggcacccct ttcatcgccg gcaagagaat
     3961 cgtgccagtg atcgagaatc acagattcac cggcagatac cgggacctgt atcctgccaa
     4021 cgagctgatc gccctgctgg aggagaaggg catcgtgttc agggatggct ccaacatcct
     4081 gccaaagctg ctggagaatg acgattctca cgccatcgac accatggtgg ccctgatccg
     4141 cagcgtgctg cagatgcgga actccaatgc cgccacaggc gaggactata tcaacagccc
     4201 cgtgcgcgat ctgaatggcg tgtgcttcga ctcccggttt cagaacccag agtggcccat
     4261 ggacgccgat gccaatggcg cctaccacat cgccctgaag ggccagctgc tgctgaatca
     4321 cctgaaggag agcaaggatc tgaagctgca gaacggcatc tccaatcagg actggctggc
     4381 ctacatccag gagctgcgca acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     4441 aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
     4501 cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4561 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4621 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4681 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     4741 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4801 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4861 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4921 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     4981 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     5041 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     5101 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     5161 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     5221 cgagctgtac aaggaattct aactagagct cgctgatcag cctcgactgt gccttctagt
     5281 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5341 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5401 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc
     5461 aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     5521 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     5581 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     5641 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     5701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5881 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     5941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6001 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     6061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6121 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     6181 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6241 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6301 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6361 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6421 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6481 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6541 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6601 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6661 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6721 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6781 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6841 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6901 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6961 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7021 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7081 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7141 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7201 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc
     7261 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7321 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7381 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7681 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
