LOCUS       V037296                 7666 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037296
VERSION     V037296
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             1648..3156
                     /label="CYP3A4(NM_017460)"
                     /note="CYP3A4(NM_017460)"
                     /gene="CYP3A4"
     polyA_signal    3233..3457
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3503..3931
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3945..4274
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4341..5135
                     /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    5309..5430
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5479..5495)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5503..5519
                     /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(5527..5557)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5572..5593
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5881..6466)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6637..7497)
                     /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(7498..7602)
                     /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 cgccaccatg gctctcatcc cagacttggc catggaaacc
     1681 tggcttctcc tggctgtcag cctggtgctc ctctatctat atggaaccca ttcacatgga
     1741 ctttttaaga agcttggaat tccagggccc acacctctgc cttttttggg aaatattttg
     1801 tcctaccata agggcttttg tatgtttgac atggaatgtc ataaaaagta tggaaaagtg
     1861 tggggctttt atgatggtca acagcctgtg ctggctatca cagatcctga catgatcaaa
     1921 acagtgctag tgaaagaatg ttattctgtc ttcacaaacc ggaggccttt tggtccagtg
     1981 ggatttatga aaagtgccat ctctatagct gaggatgaag aatggaagag attacgatca
     2041 ttgctgtctc caaccttcac cagtggaaaa ctcaaggaga tggtccctat cattgcccag
     2101 tatggagatg tgttggtgag aaatctgagg cgggaagcag agacaggcaa gcctgtcacc
     2161 ttgaaagacg tctttggggc ctacagcatg gatgtgatca ctagcacatc atttggagtg
     2221 aacatcgact ctctcaacaa tccacaagac ccctttgtgg aaaacaccaa gaagctttta
     2281 agatttgatt ttttggatcc attctttctc tcaataacag tctttccatt cctcatccca
     2341 attcttgaag tattaaatat ctgtgtgttt ccaagagaag ttacaaattt tttaagaaaa
     2401 tctgtaaaaa ggatgaaaga aagtcgcctc gaagatacac aaaagcaccg agtggatttc
     2461 cttcagctga tgattgactc tcagaattca aaagaaactg agtcccacaa agctctgtcc
     2521 gatctggagc tcgtggccca atcaattatc tttatttttg ctggctatga aaccacgagc
     2581 agtgttctct ccttcattat gtatgaactg gccactcacc ctgatgtcca gcagaaactg
     2641 caggaggaaa ttgatgcagt tttacccaat aaggcaccac ccacctatga tactgtgcta
     2701 cagatggagt atcttgacat ggtggtgaat gaaacgctca gattattccc aattgctatg
     2761 agacttgaga gggtctgcaa aaaagatgtt gagatcaatg ggatgttcat tcccaaaggg
     2821 gtggtggtga tgattccaag ctatgctctt caccgtgacc caaagtactg gacagagcct
     2881 gagaagttcc tccctgaaag attcagcaag aagaacaagg acaacataga tccttacata
     2941 tacacaccct ttggaagtgg acccagaaac tgcattggca tgaggtttgc tctcatgaac
     3001 atgaaacttg ctctaatcag agtccttcag aacttctcct tcaaaccttg taaagaaaca
     3061 cagatccccc tgaaattaag cttaggagga cttcttcaac cagaaaaacc cgttgttcta
     3121 aaggttgagt caagggatgg caccgtaagt ggagcctgag gatcctgata tccagcacag
     3181 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3241 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3301 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3361 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3421 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3481 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3541 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3601 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3661 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3721 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3781 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3841 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3901 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3961 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4021 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4081 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4141 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4201 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4261 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4321 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4381 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4441 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4501 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4561 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4621 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4681 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4741 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4801 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4861 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4921 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4981 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5041 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5101 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5161 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5221 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5281 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5341 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5401 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5461 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5521 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5581 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5641 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6061 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6301 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6421 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6481 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6541 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6601 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6661 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6721 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6781 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6841 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6901 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6961 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7021 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7081 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7141 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7201 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7261 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7321 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7381 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7441 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7501 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7561 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7621 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
