LOCUS       V042014                 8073 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042014
VERSION     V042014
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..2876
                     /label="Exd2(NM_133798)"
                     /note="Exd2(NM_133798)"
                     /gene="Exd2"
     CDS             2916..3635
                     /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"
     polyA_signal    3673..3897
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3943..4371
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4385..4714
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4781..5575
                     /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    5749..5870
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5919..5935)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5943..5959
                     /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(5967..5997)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6012..6033
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6321..6906)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7077..7937)
                     /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(7938..8042)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatgt ctagacaaaa tttagtggct ttgacagtga
      961 ccacccttct gggtgtggcc atggggggat ttgtcctgtg gaaaggcatc cagcgtagat
     1021 ggagtaagac aagcagagtg atgcagcagc agcctcagca gccacagcag ccgcagcagc
     1081 cgcagccaca gccacagcca cagccacagc cacagccaga gcatccacag ccacagcagc
     1141 aagtgcctgg tgggagggag tggccccctc cagaagatga ccagctaccc tttggtgccc
     1201 tgagagcacc cagagcctca tgggaggaga ggatactcca ggcagaggtg gtgacggtat
     1261 ctcaggaggc agagtggaat caaatccagc cctttcttaa aagagagttg gaagattttc
     1321 ctgtacttgg aattgactgt gaatgggtga atttggaagg caaagccagt cctttgtcac
     1381 tcctacagat ggcttcccca agtggcttct gtgcactggt tcgcttgccc aggctgatat
     1441 atggaggcag aacactaccc aggacgcttt tggatatatt ggcagatggt gctatcttaa
     1501 aagtaggagt gggatgttca gaagatgcca acaagcttct acaggattat ggtctcattg
     1561 tcaggggatg tctggacctt cgctacctag ccatgaagca gggaaacaat atactctgta
     1621 atgggcttag cctgaaatca cttgctgaga ctattttgaa cttcccactt gacaagtccc
     1681 ttctacttcg ttgcagcaac tgggatgctg agaatcttac tgaagaccag gtaacttatg
     1741 ctgccaggga tgcgcagatt tcagtggctc tctttctcca tcttcttgga taccctttct
     1801 ctagagattc ctatgaagag gaaagtacgg accaaatcaa ctggcaaaaa gccttggaga
     1861 gatgcaggaa tatggtagac atcccattcc gaagcaaagg acttggccgg ctggttgaag
     1921 aggttaatgg ggaagcactg gagtcgcagc tgaagccgag aaataggaag gctaagactg
     1981 atagaatggt gccaggcaac aaccaaggga gagaccccag gaagcataag aggaagcccc
     2041 tgggggtggg ctattctgcc agaaaatccc ctctctatga taactgcttc ctccaagctc
     2101 ctgatggaca gcccttgtgc acctgtgacc gcaggaaagc tcagtggtac ctggacaaag
     2161 gcattggaga gctggtgagt aaagaacctt ttgtggtcag gctacagttt gaacctgcag
     2221 gaagacctga gtccccagga gattattact tgatggttaa agagaatctg tgtgttgtct
     2281 gtggcaagac agatacctac atccggaaaa atataattcc acatgagtac cgcaagcatt
     2341 tccctattga gatgaaggac cacaactccc atgatgtgct gttgctctgc acctcctgcc
     2401 atgccatctc taactactat gacaaccatc tgaagcaaca gctggccaag gagttccagg
     2461 ctcccattgg ctcagaggag ggcctacgcc tgctggaaga cttagagcgg agacaggtgc
     2521 gttctggagc cagagccctg ctcaacgcag agagcctacc tgctcatcgg aaagaagagc
     2581 tgctgcacgc actcagggag ttttacaaca cagatattat tacagaggag atgcttcacg
     2641 aggccgccag cctggagacc aggatataca atgaaagtta tattcctcat gggctaaagg
     2701 tggtacagcg tcacaccgag ggtgggctgc gctccctcat gcagctggag agccgctggc
     2761 gtcagcactt cctggactcc atgcagccta agcacctgcc ccagcagtgg tcagtggacc
     2821 acaaccatca gaagctgctc cgaaagtacg gggatgacct tcccatcaag ctgtcaacct
     2881 cgagtggcgg tggaggaagc ggcggaggtg gatctatggt gagcaagggc gaggagctgt
     2941 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3001 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     3061 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     3121 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3181 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3241 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3301 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3361 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3421 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3481 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     3541 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3601 ggatcactct cggcatggac gagctgtaca agtagtctag agggcccgtt taaacccgct
     3661 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3721 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3781 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3841 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3901 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3961 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4021 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4081 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4141 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4201 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4261 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4321 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4381 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4441 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4501 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4561 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4621 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4681 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4741 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4801 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4861 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4921 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4981 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5041 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5101 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5161 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5221 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5281 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5341 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5401 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5461 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5521 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5581 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5641 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5701 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5761 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5821 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5881 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5941 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6001 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6061 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6121 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6841 accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6901 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6961 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7021 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7081 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7141 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7201 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7261 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7321 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7381 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7441 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7501 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7561 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7621 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7681 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7741 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7801 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7861 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7921 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7981 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8041 cgcacatttc cccgaaaagt gccacctgac gtc
//
