LOCUS       .                       7997 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        67..171
                     /label="AmpR promoter"
     CDS             172..1029
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      1203..1791
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2079..2100
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2115..2145
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2153..2169
                     /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)."
     primer_bind     2177..2193
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2214..2232
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2269..2285
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             4223..4246
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4253..4276
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4286..4309
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             6236..6952
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     polyA_signal    complement(6961..7095)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(7397..7415)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7425..7441)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      7583..8038
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
       61 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      121 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      181 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      241 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      301 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      361 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      421 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      481 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      541 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      601 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      661 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      721 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      781 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      841 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      901 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      961 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1021 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1081 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1141 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1201 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1261 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1321 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     1381 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1441 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1501 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1561 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1621 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1681 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1741 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1801 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     1861 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     1921 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     1981 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2041 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     2101 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     2161 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     2221 cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt
     2281 ggatctgctg ggcttgctga aggtaggggg tcaagaacca gaggtgtaaa gtacttgagt
     2341 aattttactt gattactgta cttaagtatt atttttgggg atttttactt tacttgagta
     2401 caattaaaaa tcaatacttt tacttttact taattacatt tttttagaaa aaaaagtact
     2461 ttttactcct tacaatttta tttacagtca aaaagtactt attttttgga gatcacttgg
     2521 gcccggctcg agttgaccaa agcttaaatc agttgtgtta aataagagac attcaaaata
     2581 aatgtaaatg agctctccaa atcagcagac ttaacattct ttaaaatgat tgattcaata
     2641 gtgataaaaa tcaggcatag ccagttgtaa ctttagataa attacagaaa atgtcaaata
     2701 cagagaaccg attctttttt atgatacatc caagcacaca tttaacacaa tccaggcaaa
     2761 ccccgaattt cacagtcaca agcactgttt gtacaagagc tttgcctaag gacacacagt
     2821 ctctataagt ccaggtcgtt ggtttcactc ttattttaaa catgtgacat ttttcctgcc
     2881 atcctgtctt aggctgctgt ttgcttcatt ccatgtcaca ttaatttcct cagtagcacc
     2941 ttttacacac acagccaatc ttttccagaa aattcaattg ctttgaaaag ataatgtgtg
     3001 aacaaatcca tttagaaaag gaaaattaag aatttgtaaa atcatctgta aattgttggc
     3061 attcttctgt atatgaacat cacatcattt acaggtaaag gtctggtcat taattatatg
     3121 acaatttact ggtattattt tgtgaaaggg gctattttca atgcattcat ccatcctttt
     3181 catccctcaa atctctcatt cacgtccccc tccccatctg cacactttat ctcattttcc
     3241 accctgctgg aatctgagca cttgtgcagt tatcagggct cctgtattta ggaggctctg
     3301 ggtgtccatg taggggacga acagaaacac tgcagacctt tatagaagaa caaatgataa
     3361 gagtcctcat acataaagac tccattagaa acgtcagtga cccaggagcc cagaccaaca
     3421 gcaaagcaga cagtgaacat ggtgagtaga caaagctata cttttttggt tttgaatata
     3481 atattaatgt gaaaataaaa agcgtctata tgaagttaaa ctaatcatta aatagtgttt
     3541 gtttgttgat attaaatatt agaagcatca ttttatgcat ttgtatgttg tgatactatc
     3601 ttttaatgag tatttttact taggtacttt tttctgtaca ctgtaaaacg caaaaaagtt
     3661 aaggtaactc aaatcatttg agaaaaccga ttgtaacaag ccatctgaag ttcaaaaact
     3721 aatctaaatg agtactgtga acttaatcta tttgagtaaa agaagcaatt tgagcacagt
     3781 aaaacccagt aaatgaagag aactcaacca actgagtact gtaaaaccca ataagatgag
     3841 gcagctcaca ccgtttgagg aaaccgattg ctacaaacca tttgagttaa aaaaaaagaa
     3901 tcaatatgag tactgtgaac ttactccatt taagttgaag taatgaggta attaattaac
     3961 tcattaccct caacactgag ttcaaaactc ttttcaaatg agtagaatta attttcagcc
     4021 aattttgagt taactatact catttcattt gataaagttg actattgggt tttacagtgt
     4081 ataaatgcta taaattgagt ttccaaataa ggcttctctt tttttttact cacaggctag
     4141 taaaaaagcc gcggccaaga gggggaaaac tgctcaaaga ggctcttcca atgtcttctc
     4201 cctcgaggga tccgccacca tggactataa agacgatgac gataagtcga tggattacaa
     4261 agacgatgat gacaaaagca gcatggatta caaggatgac gacgataaga attcaaggcc
     4321 tgtgcagaag aagagcttcc agcagagtct ggaggacaat aaagagaaga tgaaggagaa
     4381 gcggaaccgg cggctgagcg cggcggcttc ggcccggaga tcagtgcagc tccggggagc
     4441 agctgcggtg aagccgttgg tgctgaagag cgttcaggcc aacaataaag ctctggccgt
     4501 ggctctgcag gcggagcggg agaaggttcg acaggcgcag ggagtcattc tgcagctgaa
     4561 gaaggagaga caaacactgc tgttccacct gctgatgctg aagaggacac tgagagagag
     4621 acccgcgctc atcacaccac aggaggtttc tcctcctgaa gagacgagtc ccccgagacc
     4681 agctcagcct gtcagtccac ggcctgtgga tgtgtgtgat gatgtccagc agcttccgtc
     4741 agcagaggca cgtctctctg cagatgctcg atccgtgtct ttgcctccta cagtgggaat
     4801 gaggaggaaa cgctgctctg ggatgaagcg gagacgcagc tctcgcttta actcaaaccc
     4861 tgagcagatg gaggagagtg gagcccagcc cagtgtagag gacctacatc aacatcagca
     4921 ggctgaagag gcccgtgtgc agccggtgga gcctgacgtc cagctctgca ttgctgaagc
     4981 tccagcgcgc aagaagagga gcgcacagcc taaagcggag cgaggccgca aggcggatcg
     5041 ggccccgctg aagaagccct gggagaacca cagaccccga gcccgatcca agagcagaga
     5101 gcgaccacag agccgagcag gagccacgct gaccccagtc gaccgcctca actcctcact
     5161 gggaggaaac gacacctttg atttcgactg cgaggaggcc gttcacctca caccgttcag
     5221 agacaaacac aaacacaccg agacagaacc agaaccgcac gctgctgcag ctgatgggat
     5281 ggagaacagc agcccaatgg aggagcagga tgaggagaag gaagaggagg aggagacgcc
     5341 gtacatcccc aacaggagaa tccggagagc ctgcagtcct gcagccagac gagcccgatc
     5401 caaacgccac tccaacaaca tgcagacaca cagcacaggg aaaaccagca gcccaatgga
     5461 ggttgaggaa gaggaggaag agcaggagga agaggaggag acgccgtaca tccccaacag
     5521 gagaatccgg agagcctgca gtcctgcagc cagacgagcc cgatccaaac gccgctccaa
     5581 caacatgcag acacacagca caggaagctg ctctgctgct gcagtgatgg aggagcagga
     5641 cacagcagat cttctgttca cgtctccagt ctgtgatgct gaggagaaga ggagagcatc
     5701 tgacctgcag agcggcccac cacagctgag ctcatctcca gcagagtctc tggtgtccat
     5761 gatgagtcct gcagagccgg agctcctgct gctccagagc ccagtgtttg agttccagca
     5821 gaaccaggag aaccagcagc cacagccggc ccgcaggagg acaggcgggc tggtgcttcg
     5881 ctcctttctg ggcttggctt tgagtgatgt gagtaatgtg gtgtgtccag cagcgcgtca
     5941 gacgtccagc agcgcaggac acacaccaca gagctgcaga cgccagcgcc gagccgccag
     6001 cgcagtcaac tacaaagagc cctccattaa cacgaagctg cggcgcggtg ataaattcac
     6061 agacactcgt ttccttcgct ctccaatctt caagcagaaa tcttcatccc gcaggagctc
     6121 cttcaagaag atggagatct acaacgaggc cttcgtcggc tgcagaccta gggctactaa
     6181 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggacctg gatccatggt
     6241 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     6301 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     6361 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     6421 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     6481 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     6541 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     6601 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     6661 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     6721 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     6781 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     6841 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     6901 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaatgat
     6961 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     7021 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     7081 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     7141 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     7201 cctctagatc agatctaata ctcaagtaca attttaatgg agtacttttt tacttttact
     7261 caagtaagat tctagccaga tacttttact tttaattgag taaaattttc cctaagtact
     7321 tgtactttca cttgagtaaa atttttgagt actttttaca cctctgtcaa gaaccatatg
     7381 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     7441 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     7501 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     7561 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     7621 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     7681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     7741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     7801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     7861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     7921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     7981 gagctgattt aacaaaa
//
