LOCUS       V035775                 7463 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035775
VERSION     V035775
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             923..2272
                     /label="TFAP2C(NM_003222)"
                     /note="TFAP2C(NM_003222)"
                     /gene="TFAP2C"
     CDS             2315..3028
                     /codon_start=1
                     /note="excitable with blue light"
                     /product="S65T variant of Aequorea victoria green
                     fluorescent protein (Heim et al., 1995)"
                     /transl_table=1
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
                     /label="GFP (S65T)"
     polyA_signal    3063..3287
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3333..3761
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3775..4104
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4171..4965
                     /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    5139..5260
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5309..5325)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5333..5349
                     /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(5357..5387)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5402..5423
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5711..6296)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6467..7327)
                     /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(7328..7432)
                     /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 gtttaaactt aagcttgcca ccatgttgtg gaaaataacc gataatgtca agtacgaaga
      961 ggactgcgag gatcgccacg acgggagcag caatgggaat ccgcgggtcc cccacctctc
     1021 ctccgccggg cagcacctct acagccccgc gccacccctc tcccacactg gagtcgccga
     1081 atatcagccg ccaccctact ttccccctcc ctaccagcag ctggcctact cccagtcggc
     1141 cgacccctac tcgcatctgg gggaagcgta cgccgccgcc atcaaccccc tgcaccagcc
     1201 ggcgcccaca ggcagccagc agcaggcctg gcccggccgc cagagccagg agggagcggg
     1261 gctgccctcg caccacgggc gcccggccgg cctactgccc cacctctccg ggctggaggc
     1321 gggcgcggtg agcgcccgca gggatgccta ccgccgctcc gacctgctgc tgccccacgc
     1381 acacgccctg gatgccgcgg gcctggccga gaacctgggg ctccacgaca tgcctcacca
     1441 gatggacgag gtgcagaatg tcgacgacca gcacctgttg ctgcacgatc agacagtcat
     1501 tcgcaaaggt cccatttcca tgaccaagaa ccctctgaac ctcccctgtc agaaggagct
     1561 ggtgggggcc gtaatgaacc ccactgaggt cttctgctca gtccctggaa gattgtcgct
     1621 cctcagctct acgtctaaat acaaagtgac agtggctgaa gtacagaggc gactgtcccc
     1681 acctgaatgc ttaaatgcct cgttactggg aggtgttctc agaagagcca aatcgaaaaa
     1741 tggaggccgg tccttgcggg agaagttgga caagattggg ttgaatcttc cggccgggag
     1801 gcggaaagcc gctcatgtga ctctcctgac atccttagta gaaggtgaag ctgttcattt
     1861 ggctagggac tttgcctatg tctgtgaagc cgaatttcct agtaaaccag tggcagaata
     1921 tttaaccaga cctcatcttg gaggacgaaa tgagatggca gctaggaaga acatgctatt
     1981 ggcggcccag caactgtgta aagaattcac agaacttctc agccaagacc ggacacccca
     2041 tgggaccagc aggctcgccc cagtcttgga gacgaacata cagaactgct tgtctcattt
     2101 cagcctgatt acccacgggt ttggcagcca ggccatctgt gccgcggtgt ctgccctgca
     2161 gaactacatc aaagaagccc tgattgtcat agacaaatcc tacatgaacc ctggagacca
     2221 gagtccagct gattctaaca aaaccctgga gaaaatggag aaacacagga aaggtggagg
     2281 cggatcagga ggcggtggat ctggatccgc caccatgagt aaaggagaag aacttttcac
     2341 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     2401 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
     2461 tactggaaaa ctacctgttc catggccaac acttgtcact actttcactt atggtgttca
     2521 atgcttttca agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc
     2581 cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg
     2641 tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga
     2701 ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa
     2761 tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca
     2821 caacattgaa gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg
     2881 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     2941 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     3001 tacacatggc atggatgaac tatacaaata ggggcccgtt taaacccgct gatcagcctc
     3061 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3121 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3181 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3241 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3301 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3481 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3661 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3721 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3781 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3841 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4021 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4081 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4141 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4201 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4261 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4321 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4381 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4441 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4501 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4561 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4621 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4681 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4741 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4801 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4861 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4921 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4981 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5041 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5101 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5221 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5281 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5521 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6121 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6241 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6481 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6541 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6601 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6661 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6721 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6781 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6841 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6901 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6961 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7021 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7081 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7141 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7201 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7261 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7321 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7381 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7441 cccgaaaagt gccacctgac gtc
//
