LOCUS       V045422                 8023 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045422
VERSION     V045422
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             964..3513
                     /label="Tgfbr3(NM_011578)"
                     /note="Tgfbr3(NM_011578)"
                     /gene="Tgfbr3"
     CDS             3520..3585
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3623..3847
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3893..4321
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4335..4664
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4731..5525
                     /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    5699..5820
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5869..5885)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5893..5909
                     /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(5917..5947)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5962..5983
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6271..6856)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7027..7887)
                     /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(7888..7992)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcag tgacatccca ccacatggtc cctgtgtttg tcctgatgag cgcctgcctg
     1021 gccaccgcag gtccagagcc cagcacccgg tgtgaactgt caccgatcag tgcctctcat
     1081 ccagtccagg ccctgatgga gagcttcacc gttctgtctg gctgtgccag cagaggcacc
     1141 actgggctgc caagggaggt tcacatccta aacctccgca gtacagacca aggactaggc
     1201 cagccgcaga gagaggttac cctgcatctg aaccccattg cctccgtgca cactcaccac
     1261 aagcctgttg tgttcctgct caactcccca cagcccctgg tgtggcatgt gaagacagag
     1321 agactggctg ctggtgtccc cagactcttc ctggtttcgg agggttctgt ggtccagttt
     1381 tcatcaggaa acttctcctt gacagcagaa acagaggaaa ggagtttccc tcaagaaaat
     1441 gagcatctgc tacactgggc ccaaaaggaa tatggagcag tgacttcatt caccgaactt
     1501 aagatagcaa gaaatatcta tattaaagtg ggagaagatc aagtgttccc tcccacgtgt
     1561 aacataggga aaaattttct ctcgctcaat taccttgcgg agtaccttca acccaaagcc
     1621 gccgaaggtt gtgtcctggc cagtcagccc cacgagaagg aagtgcatat cattgagtta
     1681 atctccccca actccaatcc ttacagcacc ttccaggtgg atataataat tgacatacga
     1741 cctgctcgag aggatcctga ggtggtcaaa aacctcgtcc tgatcttgaa gtgcaaaaaa
     1801 tctgtcaact gggtgatcaa gtcttttgac gtcaagggaa acttgaaagt tattgctcct
     1861 gacagtattg gctttggaaa agagagtgaa cgatccatga cagtgaccaa attggtaaga
     1921 aatgacatcc cttccaccca agagaatctg atgaagtggg cattggacaa tggctacagc
     1981 ccagtgacgt catacaccat agctcctgtg gccaatagat ttcatcttcg gcttgagaac
     2041 aacgaggaga tgagagatga ggaagtccac accattcctc ctgagcttcg gatcctgctg
     2101 ggccctgacc acctgcctgc cctggacagc ccatccttcc aaggggaaat cccaaatgga
     2161 ggtttcccct tcccattccc ggatatcccc aggagaggct ggaaggaggg agaagatagg
     2221 atcccccggc caaaggaacc catcattccc agagttcaat tgcttccaga ccacagagag
     2281 ccagaagaag tgcaaggggg cgtgaatatc gccctgtcag tcaaatgtga caatgaaaag
     2341 atggtggtag ctgtagacaa agattctttc cagaccaatg gctactcggg gatggagctc
     2401 accctgttgg atccttcctg caaagccaag atgaatggta cccactttgt actggagtct
     2461 ccgctgaatg gctgtggtac tagacatagg aggtcagccc cagatggtgt ggtttactat
     2521 aactctattg tggtgcaggc tccatcccct ggggatagca gtggctggcc agacggctac
     2581 gaagatttgg agtcgggtga taatggattt cctggagaca cagatgaagg agaaactgcc
     2641 cccctgagcc gtgctggagt ggtagtgttt aactgcagct tgcggcagct gaggagtccc
     2701 agtggcttcc aggaccagct cgatggaaat gctaccttca acatggagct gtataacaca
     2761 gacctctttc tggtgccctc cccaggggtc ttctctgtgg cagagaatga gcatgtatat
     2821 gttgaggtgt ctgtcactaa ggctgaccaa gatctgggat ttgccatcca aacctgcttt
     2881 atctctccat actcaaaccc agacagaatg tctgattaca ccatcatcga gaacatctgt
     2941 ccgaaggatg actctgtgaa gttctacagc tccaagagag tgcacttccc catcccacat
     3001 gctgaagtgg acaagaagcg gttcagcttt gtgttcaagt ccgtgttcaa cacctccctg
     3061 ctcttcctgc actgcgagct gacgctgtgc tctaggaaga agggctccca gaagctgcca
     3121 aagtgtgtga ctcctgacga cgcctgcacc tctctagatg ccaccatgat ctggaccatg
     3181 atgcagaata agaagacatt caccaagccc ctggccgtgg tcctccaagt agattataaa
     3241 gaaaatgttc caaacatgaa ggagtccagt ccggttcctc ctcctccaca gattttccac
     3301 ggcctggaca cgctcaccgt gatgggcatt gcgtttgcag catttgtgat cggagcactc
     3361 ctgacggggg ccttgtggta tatctactcc cacacagggg agacggcacg aaggcagcaa
     3421 gtccctacct cgccaccagc ctcggagaac agcagcgcag cccacagcat agggagcact
     3481 cagagcactc cctgctctag cagcagcacg gccctcgagg actacaaaga ccatgacggt
     3541 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgtt
     3601 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3661 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3721 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3781 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3841 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3901 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3961 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4021 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4081 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4141 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4201 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4261 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4321 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4381 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4441 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4501 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4561 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4621 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4681 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4741 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4801 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4861 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4921 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4981 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5041 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5101 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5161 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5221 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5281 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5341 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5401 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5461 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5521 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5581 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5641 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5701 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5761 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5821 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5881 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5941 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6001 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6061 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6121 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6181 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6781 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6841 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6901 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6961 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7021 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7081 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7141 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7201 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7261 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7321 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7381 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7441 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7501 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7561 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7621 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7681 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7741 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7801 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7861 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
