LOCUS       V027385                 7570 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027385
VERSION     V027385
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..3060
                     /label="CDC25C(NM_001790)"
                     /note="CDC25C(NM_001790)"
                     /gene="CDC25C"
     polyA_signal    3137..3361
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3407..3835
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3849..4178
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4245..5039
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5213..5334
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5383..5399)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5407..5423
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /label="lac operator"
     promoter        complement(5431..5461)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5476..5497
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5785..6370)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6541..7401)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7402..7506)
                     /gene="bla"
                     /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 catgtctacg gaactcttct catccacaag agaggaagga
     1681 agctctggct caggacccag ttttaggtct aatcaaagga aaatgttaaa cctgctcctg
     1741 gagagagaca cttcctttac cgtctgtcca gatgtcccta gaactccagt gggcaaattt
     1801 cttggtgatt ctgcaaacct aagcattttg tctggaggaa ccccaaaacg ttgcctcgat
     1861 ctttcgaatc ttagcagtgg ggagataact gccactcagc ttaccacttc tgcagacctt
     1921 gatgaaactg gtcacctgga ttcttcagga cttcaggaag tgcatttagc tgggatgaat
     1981 catgaccagc acctaatgaa atgtagccca gcacagcttc tttgtagcac tccgaatggt
     2041 ttggaccgtg gccatagaaa gagagatgca atgtgtagtt catctgcaaa taaagaaaat
     2101 gacaatggaa acttggtgga cagtgaaatg aaatatttgg gcagtcccat tactactgtt
     2161 ccaaaattgg ataaaaatcc aaacctagga gaagaccagg cagaagagat ttcagatgaa
     2221 ttaatggagt tttccctgaa agatcaagaa gcaaaggtga gcagaagtgg cctatatcgc
     2281 tccccgtcga tgccagagaa cttgaacagg ccaagactga agcaggtgga aaaattcaag
     2341 gacaacacaa taccagataa agttaaaaaa aagtattttt ctggccaagg aaagctcagg
     2401 aagggcttat gtttaaagaa gacagtctct ctgtgtgaca ttactatcac tcagatgctg
     2461 gaggaagatt ctaaccaggg gcacctgatt ggtgattttt ccaaggtatg tgcgctgcca
     2521 accgtgtcag ggaaacacca agatctgaag tatgtcaacc cagaaacagt ggctgcctta
     2581 ctgtcgggga agttccaggg tctgattgag aagttttatg tcattgattg tcgctatcca
     2641 tatgagtatc tgggaggaca catccaggga gccttaaact tatatagtca ggaagaactg
     2701 tttaacttct ttctgaagaa gcccatcgtc cctttggaca cccagaagag aataatcatc
     2761 gtgttccact gtgaattctc ctcagagagg ggcccccgaa tgtgccgctg tctgcgtgaa
     2821 gaggacaggt ctctgaacca gtatcctgca ttgtactacc cagagctata tatccttaaa
     2881 ggcggctaca gagacttctt tccagaatat atggaactgt gtgaaccaca gagctactgc
     2941 cctatgcatc atcaggacca caagactgag ttgctgaggt gtcgaagcca gagcaaagtg
     3001 caggaagggg agcggcagct gcgggagcag attgcccttc tggtgaagga catgagccca
     3061 tgaggatcct gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3121 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3181 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3241 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3301 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3361 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3421 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3481 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3541 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3601 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3661 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3721 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3781 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3841 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3901 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4021 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4081 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4141 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4201 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4261 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4321 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4381 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4441 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4501 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4561 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4621 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4681 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4741 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4801 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4861 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4921 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4981 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5041 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5101 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5161 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5221 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5281 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5341 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5401 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5461 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5521 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5581 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6301 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7501 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7561 cccctatggt
//
