LOCUS       .                       5830 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        183..211
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    219..235
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
     CDS             258..911
                     /label="GST"
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     CDS             918..941
                     /label="HRV 3C site"
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             951..992
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             993..1739
                     /label="APEX2"
                     /note="soybean ascorbate peroxidase, engineered to serve as
                     a monomeric tag for electron microscopy and live-cell
                     proteomics (Lam et al., 2015)"
     promoter        2133..2237
                     /label="AmpR promoter"
     CDS             2238..3095
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3269..3857
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        4101..4178
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4179..5258
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    5274..5295
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5310..5340
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     CDS             5384..5557
                     /label="lacZα"
                     /note="LacZα fragment of β-galactosidase"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ggcaagccca
      961 tccccaaccc cctgctgggc ctggacagca ccggaaagtc ttacccaact gtgagtgctg
     1021 attaccagga cgccgttgag aaggcgaaga agaagctcag aggcttcatc gctgagaaga
     1081 gatgcgctcc tctaatgctc cgtttggcat tccactctgc tggaaccttt gacaagggca
     1141 cgaagaccgg tggacccttc ggaaccatca agcaccctgc cgaactggct cacagcgcta
     1201 acaacggtct tgacatcgct gttaggcttt tggagccact caaggcggag ttccctattt
     1261 tgagctacgc cgatttctac cagttggctg gcgttgttgc cgttgaggtc acgggtggac
     1321 ctaaggttcc attccaccct ggaagagagg acaagcctga gccaccacca gagggtcgct
     1381 tgcccgatcc cactaagggt tctgaccatt tgagagatgt gtttggcaaa gctatggggc
     1441 ttactgacca agatatcgtt gctctatctg ggggtcacac tattggagct gcacacaagg
     1501 agcgttctgg atttgagggt ccctggacct ctaatcctct tattttcgac aactcatact
     1561 tcacggagtt gttgagtggt gagaaggaag gtctccttca gctaccttct gacaaggctc
     1621 ttttgtctga ccctgtattc cgccctctcg ttgacaaata tgcagcggac gaagatgcct
     1681 tctttgctga ttacgctgag gctcaccaaa agctttccga gcttgggttt gctgatgcct
     1741 gaaagcttaa ttagctgagc ttggactcct gttgaaaggg cgaattctgc agatatccat
     1801 cacactggcg gccgctcgag cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt
     1861 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1921 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1981 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac
     2041 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2101 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2161 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2221 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2281 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2341 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2401 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2461 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2521 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2581 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2641 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2701 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2761 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2821 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2881 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2941 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3001 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3061 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3121 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3181 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3241 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3301 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3361 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3421 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3481 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3541 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3601 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3661 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3721 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3781 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3841 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3901 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3961 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4021 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4081 tttcacaccg cataaattcc gacaccatcg aatggtgcaa aacctttcgc ggtatggcat
     4141 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     4201 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     4261 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     4321 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     4381 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     4441 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     4501 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     4561 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     4621 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     4681 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     4741 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     4801 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     4861 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     4921 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     4981 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgtcaacca
     5041 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     5101 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     5161 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5221 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     5281 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5341 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga
     5401 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5461 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5521 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     5581 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     5641 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtaaccta
     5701 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     5761 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
     5821 cgttggaatt
//
