LOCUS       V043840                 8699 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043840
VERSION     V043840
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             908..4219
                     /label="Trpm8(NM_134252)"
                     /note="Trpm8(NM_134252)"
                     /gene="Trpm8"
     polyA_signal    4266..4490
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4536..4964
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4978..5307
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5374..6168
                     /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    6342..6463
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6512..6528)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6536..6552
                     /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(6560..6590)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6605..6626
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6914..7499)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7670..8530)
                     /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(8531..8635)
                     /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 cgccaccatg tccttcgagg gagccaggct cagcatgagg agccgcagaa atggtactat
      961 gggcagcacc cggaccctgt actccagtgt atctcggagc acagacgtgt cctacagtga
     1021 cagtgatttg gtgaatttta ttcaggcaaa ttttaaaaaa cgagaatgtg tcttctttac
     1081 cagagactcc aaggccatgg agaacatatg caagtgtggt tatgcccaga gccagcacat
     1141 cgaaggcacc cagatcaacc aaaatgagaa gtggaactac aaaaaacata ccaaggagtt
     1201 tccaacagac gccttcgggg acattcagtt tgagactctg gggaagaaag gcaagtactt
     1261 acgcttgtcc tgtgacaccg actctgaaac tctctacgaa ctgctgaccc agcactggca
     1321 cctcaaaaca cccaacctgg tcatttcagt gacgggtgga gccaaaaact ttgctttgaa
     1381 gccacgcatg cgcaagatct tcagcaggct gatttacatc gcacagtcta aaggtgcgtg
     1441 gattctcact ggaggcactc actacggcct gatgaagtac ataggcgagg tggtgagaga
     1501 caacaccatc agcaggaact cagaagagaa catcgtggcc attggcatcg cagcatgggg
     1561 catggtctcc aacagggaca ccctcatcag gagctgtgat gatgagggac atttttcagc
     1621 tcaatacatc atggatgact ttaccagaga ccctctatac atcctggaca acaaccatac
     1681 ccacctgctg cttgtggaca acggttgtca tggacacccc acagtggaag ccaagctccg
     1741 gaatcagctg gaaaagtaca tctctgagcg caccagtcaa gattccaact atggtggtaa
     1801 gatccccatc gtgtgttttg cccaaggagg tggaagagag actctaaaag ccatcaacac
     1861 ctctgtcaaa agcaagatcc cttgtgtggt ggtggaaggc tcggggcaga ttgctgatgt
     1921 gatcgccagc ctggtggagg tggaggatgt tttaacctct tccatggtca aagagaagct
     1981 ggtacgcttt ttaccacgca ctgtgtcccg gctgcctgaa gaggaaattg agagctggat
     2041 caaatggctc aaagaaattc ttgagagttc tcacctactc acagtaatta agatggaaga
     2101 ggctggagat gagattgtga gcaacgccat ttcctatgcg ctgtacaaag ccttcagcac
     2161 taatgagcaa gacaaggaca actggaatgg acagctgaag cttctgctgg agtggaacca
     2221 gttggacctt gccagtgatg agatcttcac caatgatcgc cgctgggagt ctgccgacct
     2281 tcaggaggtc atgttcacgg ctctcataaa ggacagaccc aagtttgtcc gcctctttct
     2341 ggagaatggc ctgaatctgc agaagtttct caccaatgaa gtcctcacag agctcttctc
     2401 cacccacttc agcaccctag tgtaccggaa tctgcagatc gccaagaact cctacaatga
     2461 cgcactcctc acctttgtct ggaagttggt ggcaaacttc cgtcgaagct tctggaaaga
     2521 ggacagaagc agcagggagg acttggatgt ggaactccat gatgcatctc tcaccacccg
     2581 gcacccgctg caagctctct tcatctgggc cattcttcag aacaagaagg aactctccaa
     2641 ggtcatttgg gagcagacca aaggctgtac tctggcagcc ttgggggcca gcaagcttct
     2701 gaagaccctg gccaaagtta agaatgatat caacgctgct ggggaatcgg aggaactggc
     2761 caatgaatat gagacccgag cagtggagtt gttcaccgag tgttacagca atgatgaaga
     2821 cttggcagaa cagctactgg tctactcctg cgaagcctgg ggtgggagca actgtctgga
     2881 gctggcagtg gaggctacag atcagcattt catcgctcag cctggggtcc agaatttcct
     2941 ttctaagcaa tggtatggag agatttcccg agacacgaag aactggaaga ttatcctgtg
     3001 tctattcatc atccccttag tgggctgtgg cctcgtatca tttaggaaga aacccattga
     3061 caagcacaag aagctgctgt ggtactatgt ggccttcttc acgtcgccct tcgtggtctt
     3121 ctcctggaac gtggtcttct acatcgcctt cctcctgctg tttgcctatg tgctgctcat
     3181 ggacttccac tcagtgccac acacccccga gctgatcctc tacgccctgg tcttcgtcct
     3241 cttctgtgat gaagtgaggc agtggtacat gaacggagtg aattatttca ccgacctatg
     3301 gaacgttatg gacaccctgg gactcttcta cttcatagcg ggtattgtat tccggctcca
     3361 ctcttctaat aaaagctcgt tgtactctgg gcgcgtcatt ttctgtctgg attacattat
     3421 attcacgcta aggctcatcc acattttcac cgtcagcagg aacttgggac ccaagattat
     3481 aatgctgcag cggatgctga tcgacgtttt cttcttcctg ttcctctttg ctgtgtggat
     3541 ggtggccttt ggcgtggcca gacaggggat cctaaggcaa aatgaacagc gctggagatg
     3601 gatcttccgc tctgtcatct atgagcccta cctggccatg tttggccagg ttcccagtga
     3661 cgtggatagt accacatatg acttctccca ctgtaccttc tcgggaaatg agtccaagcc
     3721 actgtgtgtg gagctggatg agcacaacct gccccgcttc cctgagtgga tcaccattcc
     3781 gctggtgtgc atctacatgc tctccaccaa tatccttctg gtcaacctcc tggtcgccat
     3841 gtttggctac acggtaggca ttgtacagga gaacaacgac caggtctgga aattccagcg
     3901 gtacttcctg gtgcaggagt actgcaaccg cctaaacatc cccttcccct tcgttgtctt
     3961 cgcttatttc tacatggtgg tgaagaagtg tttcaaatgc tgctgtaaag agaagaatat
     4021 ggagtctaat gcctgctgtt tcagaaatga ggacaatgag actttggcgt gggagggtgt
     4081 catgaaggag aattaccttg tcaagatcaa cacgaaagcc aacgacaact cagaggagat
     4141 gaggcatcgg tttagacaac tggactcaaa gcttaacgac ctcaaaagtc ttctgaaaga
     4201 gattgctaat aacatcaagt aactcgagtc tagagggccc gtttaaaccc gctgatcagc
     4261 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4321 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4381 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4441 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4501 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4561 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4621 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4681 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4741 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4801 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4861 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4921 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4981 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5041 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5101 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5161 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5221 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5281 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5341 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5401 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5461 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5521 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5581 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5641 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5701 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5761 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5821 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5881 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5941 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6001 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6061 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6121 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6181 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6241 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6301 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6361 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6421 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6481 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7321 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7441 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7501 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7561 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7621 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7681 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7741 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7801 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7861 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7921 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7981 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8041 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8101 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8161 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8221 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8281 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8341 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8401 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8461 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8521 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8581 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8641 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
