LOCUS       V042978                 7756 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042978
VERSION     V042978
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..3246
                     /label="Fmo2(NM_144737)"
                     /note="Fmo2(NM_144737)"
                     /gene="Fmo2"
     polyA_signal    3323..3547
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3593..4021
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4035..4364
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4431..5225
                     /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    5399..5520
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5569..5585)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5593..5609
                     /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(5617..5647)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5662..5683
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5971..6556)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6727..7587)
                     /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(7588..7692)
                     /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 catggtgaag aaggttgccg tgattggagc aggggtcagt
     1681 ggcctgatct ctctaaagtg ctgcgtggac gagggtctgg agcccacctg ctttgaaagg
     1741 actgaggata tcggaggact gtggaggttc aaagaaaacg tggaagatgg ccgagccagc
     1801 atctaccact ctgtcatcac caacactagc aaagaaatgt cctgcttcag tgacttccca
     1861 atgccggagg atttccccaa cttcctgcac aactctaaac tcctggaata cttcaggatc
     1921 tttgccaaga aatttgatct cctaaaatat attcagtttc agacaaccgt cattagtgtg
     1981 aagaaacgcc cagatttcgc atcttctggg cagtgggacg tgtatgttca gagcaatggc
     2041 aaggagcagc gtgctgtctt tgacgccgtt atggtctgca gcggccatca catccagccc
     2101 cacctgccac tcaagtcgtt tccaggtatc gagaggttcc aaggccagta tttccatagc
     2161 cgccaataca agcacccagt ggggtatgag gggaaacgca tcctagtggt tggaatagga
     2221 aactccgccg cagacattgc ctctgaactg agtaagaggg ccgcgcaggt gtttgtcagc
     2281 accagacatg gttcctgggt attgagccga atctctgaag atggctaccc ttgggacatg
     2341 gtgttccaca ctagatttag ctccatgctc cggaatgtcc tgccccgcac ggtcgtcaag
     2401 tggatgatgg aacgacagat gaatcgatgg ttcaaccatg agaattacgg cctggtgcct
     2461 cagaacaaat accttatgaa agaacctgta ttaaacgacg acctccccag ccgcctgctg
     2521 tacggagcca tcaaagtaaa aacacgggta aaggagctca cggagacagc tgtggtcttc
     2581 gaggatggca cggtggagga agacgtggat gtcattgtct ttgccacagg gtatacgttc
     2641 tcttttcctt tcctggaaga ctcgcttgtt aaagtagagg ataataaggt ctcgctgtac
     2701 aaagccatgt ttccccctca cctggagaag ccaacccttg cgtgcatagg tctcatccag
     2761 cctctgggct ccatcttccc aaccgtagag cttcaggcgc ggtgggcgac aagagttttc
     2821 aaaggcgtgt gtcgcttgcc ttcggagacg acgatgatgg cagacattgc tgaaagaaat
     2881 gagaaaagaa ttgacctgtt tggaaaaagc cagagccaga ttctgcagac caattacatt
     2941 gattacctgg atgaactcgc cttggagata ggtgcgaagc cggacttcat ctctctcctt
     3001 ttcaaagacc ctaaactggc tgtgaaactc tacttcggac cctgcaattc ctaccagtac
     3061 cgcctagttg gacctgggca gtgggaagga gccaggaatg ccatcctcac ccagaagcag
     3121 cggatcctga agccgttaaa gactcgcact ctccagacct cagctagtgc cccagtgtct
     3181 ttcctgatta aggtcctggg gctgcttgct attgttctgg ccttcttttt caagcttcat
     3241 gggttctaag gatcctgata tccagcacag tggcggccgc tcgagtctag agggcccgtt
     3301 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3361 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3421 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3481 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3541 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3601 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3661 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3721 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3781 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3841 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3901 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3961 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4021 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4141 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4201 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4261 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4321 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4381 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4441 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4501 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4561 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4621 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4681 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4741 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4801 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4861 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4921 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4981 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5041 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5101 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5161 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5221 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5281 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5341 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5401 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5461 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5521 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5581 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5641 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5701 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5761 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5821 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6481 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7741 cccgatcccc tatggt
//
