LOCUS       V029295                 8416 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029295
VERSION     V029295
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             931..3855
                     /label="Trpc5(NM_009428)"
                     /note="Trpc5(NM_009428)"
                     /gene="Trpc5"
     CDS             3862..3888
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3892..3918
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3922..3948
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3983..4207
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4253..4681
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4695..5024
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5091..5885
                     /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    6059..6180
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6229..6245)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6253..6269
                     /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(6277..6307)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6322..6343
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6631..7216)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7387..8247)
                     /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(8248..8352)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggcccagc tgtactacaa gaaggtcaat
      961 tactcaccat acagagatcg catccccctc caaatcgtga gggctgagac tgagctgtct
     1021 gctgaggaga aagccttcct tagtgctgtg gagaaggggg actatgccac agtgaagcag
     1081 gcactgcagg aggccgagat ctactacaat gtcaacatca actgcatgga tcctttgggc
     1141 aggagcgccc tgctcattgc cattgagaat gagaacttgg agatcatgga gctactgctg
     1201 aaccacagtg tgtatgtggg cgatgcatta ctctacgcca tccgcaagga ggtggtagga
     1261 gctgtggaac ttctgcttag ctacaggaag cccagtggag agaagcaggt tcccacgctg
     1321 atgatggaca cccagttctc tgagttcaca ccagacatca ctcccattat gttggcagcc
     1381 cacaccaaca actatgaaat catcaaattg cttgtccaaa agagggtcac tatcccaagg
     1441 ccccaccaga tccgctgcaa ctgtgtggag tgtgtatcca gttcggaggt agatagtcta
     1501 cgtcattcta ggtcccgact gaatatatac aaggctctgg caagcccctc gctcattgcc
     1561 ttatcaagcg aggaccccat cctcacggcc tttcggctgg gctgggaact caaggagctg
     1621 agcaaagtgg aaaatgagtt caaggctgag tacgaggaac tttcccagca atgtaagctc
     1681 tttgccaagg atctcttgga ccaagctcgg agctccagag aactggagat catcctcaac
     1741 catcgagatg accacagtga agaacttgac ccacagaagt atcatgacct ggccaagctg
     1801 aaggtggcaa tcaaatatca ccagaaagag tttgtcgctc agccaaactg ccaacaattg
     1861 ctagccacct tgtggtacga tggcttccct ggatggcgga gaaaacactg ggtagtcaag
     1921 cttctgacct gcatgaccat tggattcctg tttcctatgc tgtccatagc ctatctgatc
     1981 tcacccagaa gcaaccttgg gcttttcatc aagaaaccct ttattaagtt catctgccac
     2041 acagcatcct atctgacctt cctcttcatg cttctcctgg cttctcaaca tatcgtcaga
     2101 acagacctcc atgttcaagg acctccccca actgttgtgg aatggatgat actgccttgg
     2161 gttctaggtt tcatttgggg ggagataaag gaaatgtggg atggtggatt cacggaatac
     2221 atccatgatt ggtggaacct gatggatttt gcaatgaact ccctctacct ggcaactatt
     2281 tccttgaaga tcgtggccta tgtcaagtat aatggttctc gtccaaggga agaatgggaa
     2341 atgtggcacc cgactctgat tgcagaagca ctcttcgcaa tatccaacat tttaagttcc
     2401 ttgcgtctca tatccttgtt cacagccaac tcccatttag ggcctctgca gatctctttg
     2461 ggacgaatgc tgcttgatat actcaaattt ctctttatct actgcctagt actactggct
     2521 tttgccaacg gactgaacca gctttacttt tactatgaga ccagagctat tgatgaacct
     2581 aacaactgca aggggatccg atgtgaaaaa cagaacaacg ccttctccac gctctttgaa
     2641 acccttcagt cgctcttctg gtctgtcttt ggcctcctaa atctctatgt taccaatgtg
     2701 aaggcccgac atgagttcac tgagtttgtg ggagctacga tgtttgggac atacaacgtc
     2761 atctccctgg tagtgctgct gaatatgctc atcgccatga tgaacaactc ctaccagctc
     2821 attgccgatc atgctgatat tgagtggaag tttgcgagaa caaagctctg gatgagttac
     2881 tttgatgaag gtggcacctt accacctcct ttcaacatta ttcccagccc caaatcattt
     2941 ctctatcttg gcaactggtt caacaacacc ttctgtccca aaagagaccc tgatggaaga
     3001 cgaaggaggc acaacttgag aagcttcaca gaacgtcatg ctgacagcct gatacaaaat
     3061 caacattatc aggaagttat cagaaattta gtcaagagat atgtggctgc aatgataaga
     3121 aactccaaaa caaatgaggg gctaacagaa gaaaatttta aggaattaaa gcaggacatt
     3181 tccagctttc gatatgaagt gcttgacctc ttgggaaaca ggaaacatcc aagaagaagc
     3241 ctctccacca gcagtgctga tttctctcaa agggatgata ccaatgacgg cagtggtggg
     3301 gctcgggcca agtctaagag tgtctctttc aatgtaggct gcaagaaaaa ggcctgtcac
     3361 ggtgcaccgc tcatcagaac cgtgccaaga gctagtggcg cccaaggaaa gccaaaatcc
     3421 gagtcgtcga gcaaacggtc cttcatgggt ccgtctttca agaaattggg tctcttcttc
     3481 tccaagttca atggtcaaac atctgaacct acttcggagc caatgtacac aatctctgat
     3541 ggcatcgcac agcagcattg tatgtggcag gacatccgct attctcagat ggagaaaggg
     3601 aaggcagagg cctgttctca aagtcagatg aaccttggcg aggtagagct aggtgaaatc
     3661 cggggggctg ctgctcggag cagtgagtgc cccctagcct gttccagctc ccttcactgt
     3721 gcatccggca tctgctcctc aaattcgaaa cttttggact cgtcagagga cgtatttgaa
     3781 acttggggag aggcttgtga cttgctcatg cacaaatggg gtgatggaca ggaagaacaa
     3841 gttacaactc ggctcctcga gtacccatac gacgtacctg attatgcaac ttatccttat
     3901 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcatg agggcccgtt
     3961 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4021 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4081 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4141 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4201 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4261 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4321 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4381 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4441 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4501 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4561 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4621 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4681 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4861 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4921 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4981 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5041 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5101 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5161 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5221 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5281 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5341 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5401 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5461 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5521 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5581 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5641 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5701 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5761 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5821 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5881 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5941 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6001 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6061 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6121 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6181 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6361 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7141 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     7201 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7261 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7321 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7381 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7441 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7501 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7561 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7621 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7681 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7741 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7801 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7861 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7921 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7981 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8041 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8101 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8161 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8221 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8281 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8341 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     8401 cccgatcccc tatggt
//
