LOCUS       .                       6450 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        582..785
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        830..848
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             874..1590
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     CDS             1918..1941
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     polyA_signal    2017..2241
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2287..2715
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2729..3058
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3125..3916
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4093..4226
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4263..4279)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4287..4303)
                     /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)."
     promoter        complement(4311..4341)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4356..4377)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4665..5250)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5424..6281)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(6282..6386)
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgcagatc tttgtgaaaa ccttaactgg taagaccatc
     1681 accctggagg tcgagcccag tgacaccatt gagaatgtca aggcaaagat ccaggacaag
     1741 gagggcatcc cccctgacca gcagaggctg atctttgcag gcaagcagct ggaagatggc
     1801 cgcaccctgt cagactacaa catccagaaa gagtccaccc tgcacctggt ccttcgcctc
     1861 agaggtggcc tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     1921 tacaaggatg acgatgacaa gtaggatcct gatatccagc acagtggcgg ccgctcgagt
     1981 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2041 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2101 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2161 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2221 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2281 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2341 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2401 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2461 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2521 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2581 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2641 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2701 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2761 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2821 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2881 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2941 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3001 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3061 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3121 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3181 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3241 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3301 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3361 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3421 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3481 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3541 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3601 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3661 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3721 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3781 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3841 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3901 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3961 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4081 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4201 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4321 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     5221 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5281 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5341 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5401 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5461 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5521 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5581 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5641 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5701 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5761 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5821 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5881 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5941 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6001 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6061 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6121 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6181 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6241 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6301 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6361 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6421 ggatcgggag atctcccgat cccctatggt
//
