LOCUS       V047647                 8128 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047647
VERSION     V047647
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..2916
                     /label="SMAD3(NM_005902)"
                     /note="SMAD3(NM_005902)"
                     /gene="SMAD3"
     CDS             2965..3621
                     /codon_start=1
                     /gene=""
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /transl_table=1
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK*"
                     /label="GST"
     polyA_signal    3695..3919
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3965..4393
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4407..4736
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4803..5597
                     /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    5771..5892
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5941..5957)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5965..5981
                     /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(5989..6019)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6034..6055
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6343..6928)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7099..7959)
                     /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(7960..8064)
                     /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 catgtcgtcc atcctgcctt tcactccccc gatcgtgaag
     1681 cgcctgctgg gctggaagaa gggcgagcag aacgggcagg aggagaaatg gtgcgagaag
     1741 gcggtcaaga gcctggtcaa gaaactcaag aagacggggc agctggacga gctggagaag
     1801 gccatcacca cgcagaacgt caacaccaag tgcatcacca tccccaggtc cctggatggc
     1861 cggttgcagg tgtcccatcg gaaggggctc cctcatgtca tctactgccg cctgtggcga
     1921 tggccagacc tgcacagcca ccacgagcta cgggccatgg agctgtgtga gttcgccttc
     1981 aatatgaaga aggacgaggt ctgcgtgaat ccctaccact accagagagt agagacacca
     2041 gttctacctc ctgtgttggt gccacgccac acagagatcc cggccgagtt ccccccactg
     2101 gacgactaca gccattccat ccccgaaaac actaacttcc ccgcaggcat cgagccccag
     2161 agcaatattc cagagacccc accccctggc tacctgagtg aagatggaga aaccagtgac
     2221 caccagatga accacagcat ggacgcaggt tctccaaacc tatccccgaa tccgatgtcc
     2281 ccagcacata ataacttgga cctgcagcca gttacctact gcgagccggc cttctggtgc
     2341 tccatctcct actacgagct gaaccagcgc gtcggggaga cattccacgc ctcgcagcca
     2401 tccatgactg tggatggctt caccgacccc tccaattcgg agcgcttctg cctagggctg
     2461 ctctccaatg tcaacaggaa tgcagcagtg gagctgacac ggagacacat cggaagaggc
     2521 gtgcggctct actacatcgg aggggaggtc ttcgcagagt gcctcagtga cagcgctatt
     2581 tttgtccagt ctcccaactg taaccagcgc tatggctggc acccggccac cgtctgcaag
     2641 atcccaccag gatgcaacct gaagatcttc aacaaccagg agttcgctgc cctcctggcc
     2701 cagtcggtca accagggctt tgaggctgtc taccagttga cccgaatgtg caccatccgc
     2761 atgagcttcg tcaaaggctg gggagcggag tacaggagac agactgtgac cagtaccccc
     2821 tgctggattg agctgcacct gaatgggcct ttgcagtggc ttgacaaggt cctcacccag
     2881 atgggctccc caagcatccg ctgttccagt gtgtctggag gagggggaag cggaggcgga
     2941 gggagtaaga attctgcaga tatcatgtcc cctatactag gttattggaa aattaagggc
     3001 cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg
     3061 tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt
     3121 cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata
     3181 cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt
     3241 tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt
     3301 aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg
     3361 ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac
     3421 ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg
     3481 ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac
     3541 ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt
     3601 ggcgaccatc ctccaaaatg aggatcctga tatccagcac agtggcggcc gctcgagtct
     3661 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3721 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3781 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3841 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3901 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3961 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4021 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4081 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4141 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4201 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4261 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4321 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4381 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4441 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4501 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4561 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4621 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4681 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4741 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4801 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4861 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4921 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4981 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5041 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5101 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5161 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5221 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5281 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5341 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5401 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5461 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5521 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5581 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5641 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5701 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5761 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5821 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5881 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5941 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6001 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6061 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6121 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6181 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6241 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6301 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6361 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6421 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6481 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6541 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6601 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6661 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6721 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6781 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6841 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6901 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6961 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7021 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7081 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7141 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7201 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7261 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7321 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7381 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7441 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7501 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7561 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7621 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7681 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7741 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7801 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7861 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7921 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7981 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8041 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8101 atcgggagat ctcccgatcc cctatggt
//
