LOCUS       V041546                 7931 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041546
VERSION     V041546
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..3424
                     /label="TRPV1(NM_080706)"
                     /note="TRPV1(NM_080706)"
                     /gene="TRPV1"
     CDS             3434..3451
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3498..3722
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3768..4196
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4210..4539
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4606..5400
                     /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    5574..5695
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5744..5760)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5768..5784
                     /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(5792..5822)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5837..5858
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6146..6731)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6902..7762)
                     /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(7763..7867)
                     /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 aagaaatgga gcagcacaga cttgggggca gctgcggacc cactccaaaa
      961 ggacacctgc ccagaccccc tggatggaga ccctaactcc aggccacctc cagccaagcc
     1021 ccagctctcc acggccaaga gccgcacccg gctctttggg aagggtgact cggaggaggc
     1081 tttcccggtg gattgccctc acgaggaagg tgagctggac tcctgcccga ccatcacagt
     1141 cagccctgtt atcaccatcc agaggccagg agacggcccc accggtgcca ggctgctgtc
     1201 ccaggactct gtcgccgcca gcaccgagaa gaccctcagg ctctatgatc gcaggagtat
     1261 ctttgaagcc gttgctcaga ataactgcca ggatctggag agcctgctgc tcttcctgca
     1321 gaagagcaag aagcacctca cagacaacga gttcaaagac cctgagacag ggaagacctg
     1381 tctgctgaaa gccatgctca acctgcacga cggacagaac accaccatcc ccctgctcct
     1441 ggagatcgcg cggcaaacgg acagcctgaa ggagcttgtc aacgccagct acacggacag
     1501 ctactacaag ggccagacag cactgcacat cgccatcgag agacgcaaca tggccctggt
     1561 gaccctcctg gtggagaacg gagcagacgt ccaggctgcg gcccatgggg acttctttaa
     1621 gaaaaccaaa gggcggcctg gattctactt cggtgaactg cccctgtccc tggccgcgtg
     1681 caccaaccag ctgggcatcg tgaagttcct gctgcagaac tcctggcaga cggccgacat
     1741 cagcgccagg gactcggtgg gcaacacggt gctgcacgcc ctggtggagg tggccgacaa
     1801 cacggccgac aacacgaagt ttgtgacgag catgtacaat gagattctga tgctgggggc
     1861 caaactgcac ccgacgctga agctggagga gctcaccaac aagaagggaa tgacgccgct
     1921 ggctctggca gctgggaccg ggaagatcgg ggtcttggcc tatattctcc agcgggagat
     1981 ccaggagccc gagtgcaggc acctgtccag gaagttcacc gagtgggcct acgggcccgt
     2041 gcactcctcg ctgtacgacc tgtcctgcat cgacacctgc gagaagaact cggtgctgga
     2101 ggtgatcgcc tacagcagca gcgagacccc taatcgccac gacatgctct tggtggagcc
     2161 gctgaaccga ctcctgcagg acaagtggga cagattcgtc aagcgcatct tctacttcaa
     2221 cttcctggtc tactgcctgt acatgatcat cttcaccatg gctgcctact acaggcccgt
     2281 ggatggcttg cctcccttta agatggaaaa aactggagac tatttccgag ttactggaga
     2341 gatcctgtct gtgttaggag gagtctactt ctttttccga gggattcagt atttcctgca
     2401 gaggcggccg tcgatgaaga ccctgtttgt ggacagctac agtgagatgc ttttctttct
     2461 gcagtcactg ttcatgctgg ccaccgtggt gctgtacttc agccacctca aggagtatgt
     2521 ggcttccatg gtattctccc tggccttggg ctggaccaac atgctctact acacccgcgg
     2581 tttccagcag atgggcatct atgccgtcat gatagagaag atgatcctga gagacctgtg
     2641 ccgtttcatg tttgtctaca tcgtcttctt gttcgggttt tccacagcgg tggtgacgct
     2701 gattgaagac gggaagaatg actccctgcc gtctgagtcc acgtcgcaca ggtggcgggg
     2761 gcctgcctgc aggccccccg atagctccta caacagcctg tactccacct gcctggagct
     2821 gttcaagttc accatcggca tgggcgacct ggagttcact gagaactatg acttcaaggc
     2881 tgtcttcatc atcctgctgc tggcctatgt aattctcacc tacatcctcc tgctcaacat
     2941 gctcatcgcc ctcatgggtg agactgtcaa caagatcgca caggagagca agaacatctg
     3001 gaagctgcag agagccatca ccatcctgga cacggagaag agcttcctta agtgcatgag
     3061 gaaggccttc cgctcaggca agctgctgca ggtggggtac acacctgatg gcaaggacga
     3121 ctaccggtgg tgcttcaggg tggacgaggt gaactggacc acctggaaca ccaacgtggg
     3181 catcatcaac gaagacccgg gcaactgtga gggcgtcaag cgcaccctga gcttctccct
     3241 gcggtcaagc agagtttcag gcagacactg gaagaacttt gccctggtcc cccttttaag
     3301 agaggcaagt gctcgagata ggcagtctgc tcagcccgag gaagtttatc tgcgacagtt
     3361 ttcagggtct ctgaagccag aggacgctga ggtcttcaag agtcctgccg cttccgggga
     3421 gaaggcggcc gcacatcacc atcatcacca ttgactcgag tctagagggc ccgtttaaac
     3481 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3541 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3601 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3661 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3721 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3781 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3841 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3901 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3961 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4021 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4081 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4141 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4441 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4561 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4621 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4681 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4741 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4801 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4861 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4921 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4981 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5041 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5101 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5161 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5221 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5281 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5341 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5401 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5461 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5521 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5581 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5641 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5701 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5761 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5821 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5881 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5941 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6661 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6721 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6781 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6841 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6901 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6961 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7021 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7081 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7141 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7201 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7261 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7321 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7381 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7441 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7501 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7561 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7621 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7681 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7741 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7801 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7861 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7921 tcccctatgg t
//
