LOCUS       V034296                 7881 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034296
VERSION     V034296
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2561
                     /label="TBX15(NM_001330677)"
                     /note="TBX15(NM_001330677)"
                     /gene="TBX15"
     CDS             2568..2633
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2657..3245
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3311..3359
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3561..3989
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4003..4332
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4399..5193
                     /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    5369..5490
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5539..5555)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5563..5579
                     /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(5587..5617)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5632..5653
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5941..6529)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6700..7560)
                     /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(7561..7665)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgag tgaaaggaga agatctgcag
      781 tcgccctgag ctcgcgagca catgccttct ccgttgaagc cttgatcggc tcaaataaaa
      841 aacggaaact gcgagactgg gaggagaagg ggctggacct gtctatggag gcgctgagcc
      901 ccgcgggccc actcggagac acggaggacg cggcggcaca cggcctggag cctcacccgg
      961 attctgagca gagcactggt tcagattctg aggtcctcac tgagcggact tcctgctcct
     1021 tcagtactca cactgacctg gcctctggtg ctgcaggccc tgtgcctgct gccatgtctt
     1081 ccatggagga gattcaggtg gagctgcaat gtgctgacct ctggaagcgg ttccatgata
     1141 ttggaactga aatgatcatc accaaagcag gcaggaggat gtttcctgcc atgagagtga
     1201 aaatcactgg cctagatcca catcagcagt actacatagc aatggacatt gtgcctgtgg
     1261 acaataaaag atacagatat gtgtatcata gctccaagtg gatggtggct ggcaatgctg
     1321 attcccctgt gcccccaaga gtttatatac accctgattc tctagcttct ggagacacct
     1381 ggatgagaca ggtggtcagt tttgacaaac tcaagcttac caacaatgag ttggatgatc
     1441 aaggacatat cattctgcac tctatgcaca aataccagcc tcgagttcat gtgattcgca
     1501 aagacttcag cagtgacctt tcacccacta agcctgttcc tgttggggat ggggtgaaaa
     1561 cgttcaactt tcctgagact gtgttcacca cagttacggc ctatcagaat cagcagatta
     1621 ccagattaaa aattgaccga aacccttttg ctaaaggatt cagagattct gggagaaaca
     1681 gaactggact tgaagccatc atggagacat atgcattctg gagacctcct gtgcgcacac
     1741 tcaccttcga agacttcacc accatgcaga agcagcaagg aggcagcaca ggcacttccc
     1801 caaccacctc cagcactggg acaccatccc cttcggcttc ttctcatctt ttatctccat
     1861 cctgttctcc tccaactttt catctggccc ccaacacttt caatgtgggc tgccgagaaa
     1921 gccagctgtg taatctaaac ctctctgatt atccaccatg tgcccgaagc aacatggctg
     1981 ccttgcagag ctacccaggg ctgagtgaca gtggctacaa caggcttcag agtggcacca
     2041 cttcagccac tcagccctct gaaaccttca tgcctcagag gactccatcc ctgatctcag
     2101 gaataccaac tcctccctcg ttgcctggca acagcaagat ggaagcctac ggtggccagc
     2161 tggggtcctt tcccacttcc cagtttcagt atgtcatgca ggcaggcaat gctgcctcca
     2221 gctcctcatc accacacatg ttcgggggca gccacatgca gcagagctcc tacaatgcct
     2281 tctcccttca caacccttac aacctgtatg gatacaattt ccccacttcc cctaggctag
     2341 ctgcaagccc ggaaaaactg agcgcctctc aaagcacttt actctgttct tctccttcca
     2401 acggggcctt tggagagagg cagtacctgc cgtcagggat ggagcacagc atgcacatga
     2461 ttagcccttc acccaataac caacaggcaa ccaacacttg tgatggccgg cagtatgggg
     2521 cagttccagg ctcctcctcc cagatgtccg tgcacatggt tggatccgac tacaaagacc
     2581 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     2641 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2701 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2761 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2821 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2881 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2941 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3001 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     3061 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3121 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3181 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3241 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3301 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     3361 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3421 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3481 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     3541 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3601 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3661 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     3721 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3781 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3841 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3901 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3961 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4141 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4201 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4261 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4321 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4381 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4441 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4501 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4561 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4621 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4681 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4741 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4801 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4861 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4921 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4981 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5041 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5101 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5161 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     5221 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5281 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5341 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5401 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5461 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5521 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5581 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5641 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5701 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6121 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6361 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6481 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6541 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6601 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6661 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6961 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7141 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7441 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7561 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7621 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7681 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     7741 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7801 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7861 tgcatgaaga atctgcttag g
//
