LOCUS       V039666                 8959 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039666
VERSION     V039666
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1030..4479
                     /label="CCAR1(NM_018237)"
                     /note="CCAR1(NM_018237)"
                     /gene="CCAR1"
     polyA_signal    4559..4783
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4829..5257
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5271..5600
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5667..6461
                     /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    6635..6756
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6805..6821)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6829..6845
                     /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(6853..6883)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6898..6919
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7207..7792)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7963..8823)
                     /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(8824..8928)
                     /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 aagccaccat ggcggcgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
     1021 gaaaagctta tggctcaatt tggaggacag aagaatccgc catgggctac tcagtttaca
     1081 gccactgcag tatcacagcc agctgcactg ggtgttcaac agccatcact ccttggagca
     1141 tctcctacca tttatacaca gcaaactgca ttggcagcag caggccttac cacacaaact
     1201 ccagcaaact atcagttaac acaaactgct gcattgcagc aacaagccgc agctgcagca
     1261 gctgcattac aacagcaata ttcacaacct cagcaggccc tgtatagtgt gcaacaacag
     1321 ttacagcaac cccagcaaac cctcttaaca cagccagctg ttgcactgcc tacaagcctt
     1381 agcctgtcta ctcctcagcc aacagcacaa ataactgtat catatccaac accaaggtcc
     1441 agtcaacagc aaacccagcc tcagaagcag cgtgttttca caggggtggt tacaaaacta
     1501 catgatacgt ttggatttgt ggatgaagat gtattctttc agcttagtgc tgtcaaaggg
     1561 aaaacccccc aagtaggtga cagagtattg gttgaagcta cttataatcc taatatgcct
     1621 tttaaatgga atgcacagag aattcaaaca ctaccaaatc agaatcagtc gcaaacccag
     1681 ccattactga agactcctcc tgctgtactt cagccaattg caccacagac aacatttggt
     1741 gttcagactc agccccagcc ccagtcactg ctgcaggcac agatttcagc agcttctatt
     1801 acaccactat tgcagactca accacagccc ttattacagc agcctcagca aaaagctggt
     1861 ttattgcagc ctcctgttcg tatagtttca cagccacaac cggcacgacg attagatccc
     1921 ccatcccgat tttcaggaag aaatgacaga ggggatcaag tgcctaacag aaaagatgat
     1981 cgaagtcgtg agagagagag agaaagacgt agatcgagag aaagatcacc tcagaggaaa
     2041 cgttcccggg aaagatctcc acgaagagag cgagagcgat cacctcggag agttcgacgt
     2101 gttgttccac gttacacagt tcagttttca aagttttctt tagattgtcc cagttgtgac
     2161 atgatggaac taaggcgccg ttatcaaaat ttgtatatac ctagtgactt ttttgatgct
     2221 caatttacat gggtggatgc tttccctttg tcaagaccat ttcagctggg aaattactgc
     2281 aatttttatg taatgcacag agaagtagag tccttagaaa aaaatatggc cattcttgat
     2341 ccaccagatg ctgaccactt atacagtgca aaggtaatgc tgatggctag ccctagtatg
     2401 gaagatttat atcataagtc atgtgctctt gctgaggacc cacaagaact tcgagatgga
     2461 ttccaacatc ctgctagact tgttaagttt ttagtgggca tgaaaggcaa ggatgaagct
     2521 atggccattg gaggccactg gtctccttcg ttggatggac cagacccaga aaaagatccc
     2581 tctgtgttga ttaagactgc tattcgttgt tgtaaggctc tgacaggcat tgatctaagt
     2641 gtgtgcacac aatggtaccg ttttgcagag attcgctacc atcgccctga ggagacccac
     2701 aaggggcgta cagttccagc tcatgtggag acagtggttt tatttttccc ggatgtttgg
     2761 cattgccttc ccacccgctc agagtgggaa accctctccc gaggatacaa gcagcagctg
     2821 gtcgagaagc ttcagggtga acgcaaggag gctgatggag aacaggatga agaagagaag
     2881 gatgatggtg aagctaaaga aatttctaca cctacccatt ggtctaaact tgatccaaag
     2941 acaatgaagg taaatgacct ccgaaaagaa ttagaaagtc gagctcttag ttccaaagga
     3001 ttaaaatccc agttaatagc ccgattgaca aaacagctta aagtagagga acaaaaagaa
     3061 gaacagaagg agttagagaa atctgaaaaa gaagaggatg aggatgatga taggaaatct
     3121 gaagacgata aagaggaaga agaaaggaaa cgtcaagagg aaatagaacg ccagcgtcga
     3181 gaaagaagat atattttgcc tgatgaaccg gccatcattg tacatccaaa ttgggctgca
     3241 aaaagtggca agtttgattg tagcatcatg tctttgagtg tcctattgga ctacagatta
     3301 gaggataata aagaacattc atttgaggtt tcattgtttg cggaactttt caacgaaatg
     3361 cttcaaagag attttggtgt ccgtatatac aaatcattac tgtctcttcc tgagaaagag
     3421 gacaaaaaag aaaaggataa aaaaagcaaa aaagatgaga gaaaagataa aaaagaagaa
     3481 agagatgatg aaactgatga accaaaaccc aaacggagaa aatcaggcga tgataaagat
     3541 aaaaaagaag atagagatga aaggaagaaa gaagataaaa gaaaagatga ttctaaagat
     3601 gatgatgaaa ctgaagaaga taacaatcaa gatgaatatg accctatgga agcagaagaa
     3661 gctgaggatg aagaagatga tagggatgag gaagaaatga ccaaacgaga tgacaaaaga
     3721 gatatcaaca gatactgcaa ggagaggccc tctaaagata aggaaaaaga aaagactcaa
     3781 atgatcacaa ttaacagaga tctgttaatg gcttttgttt attttgatca aagtcattgt
     3841 ggttaccttc ttgaaaagga tttggaagaa atactttata ctcttggact acatctttct
     3901 cgggctcagg taaagaagct tcttaataaa gtagtgctcc gtgaatcttg cttttaccgg
     3961 aaattaacag acacctcaaa agatgaagag aaccatgaag agtctgagtc attgcaggaa
     4021 gatatgctag gaaacagatt attacttcca acaccaacag taaagcagga atcaaaggat
     4081 gtggaagaaa atgttggcct cattgtgtac aatggtgcaa tggtagatgt aggaagcctc
     4141 ttgcaaaaat tggaaaagag cgaaaaagta agagctgagg tagaacagaa gctgcagtta
     4201 ctagaagaaa aaacagatga agatgaaaaa accatattaa atttggagaa ttccaacaaa
     4261 agcctctctg gtgaactcag agaagttaaa aaggacctta gtcagttaca agaaaactta
     4321 aagatttcgg aaaacatgaa tttacaattt gaaaaccaaa tgaataagac aatcagaaac
     4381 ttatctacgg taatggatga aatccacact gttctcaaga aggataatgt aaagaatgaa
     4441 gacaaagatc aaaaatccaa ggagaatggt gccagtgtat gagaattctg cagatatcca
     4501 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     4561 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4621 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4681 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4741 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4801 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4861 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4921 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4981 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5041 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5101 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5161 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5221 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     5281 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5341 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5401 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5461 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5521 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5581 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     5641 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5701 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5761 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5821 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5881 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5941 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     6001 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     6061 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6121 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6181 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6241 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6301 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6361 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6421 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6481 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6541 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6601 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6661 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6721 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6781 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7741 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8941 aaaagtgcca cctgacgtc
//
