LOCUS       V032692                 7882 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032692
VERSION     V032692
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..3318
                     /label="TLR6(NM_001394553)"
                     /note="TLR6(NM_001394553)"
                     /gene="TLR6"
     CDS             3325..3414
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3449..3673
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3719..4147
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4161..4490
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4557..5351
                     /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    5525..5646
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5695..5711)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5719..5735
                     /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(5743..5773)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5788..5809
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6097..6682)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6853..7713)
                     /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(7714..7818)
                     /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 atgaccaaag acaaagaacc tattgttaaa
      961 agcttccatt ttgtttgcct tatgatcata atagttggaa ccagaatcca gttctccgac
     1021 ggaaatgaat ttgcagtaga caagtcaaaa agaggtctta ttcatgttcc aaaagaccta
     1081 ccgctgaaaa ccaaagtctt agatatgtct cagaactaca tcgctgagct tcaggtctct
     1141 gacatgagct ttctatcaga gttgacagtt ttgagacttt cccataacag aatccagcta
     1201 cttgatttaa gtgttttcaa gttcaaccag gatttagaat atttggattt atctcataat
     1261 cagttgcaaa agatatcctg ccatcctatt gtgagtttca ggcatttaga tctctcattc
     1321 aatgatttca aggccctgcc catctgtaag gaatttggca acttatcaca actgaatttc
     1381 ttgggattga gtgctatgaa gctgcaaaaa ttagatttgc tgccaattgc tcacttgcat
     1441 ctaagttata tccttctgga tttaagaaat tattatataa aagaaaatga gacagaaagt
     1501 ctacaaattc tgaatgcaaa aacccttcac cttgtttttc acccaactag tttattcgct
     1561 atccaagtga acatatcagt taatacttta gggtgcttac aactgactaa tattaaattg
     1621 aatgatgaca actgtcaagt tttcattaaa tttttatcag aactcaccag aggttcaacc
     1681 ttactgaatt ttaccctcaa ccacatagaa acgacttgga aatgcctggt cagagtcttt
     1741 caatttcttt ggcccaaacc tgtggaatat ctcaatattt acaatttaac aataattgaa
     1801 agcattcgtg aagaagattt tacttattct aaaacgacat tgaaagcatt gacaatagaa
     1861 catatcacga accaagtttt tctgttttca cagacagctt tgtacaccgt gttttctgag
     1921 atgaacatta tgatgttaac catttcagat acacctttta tacacatgct gtgtcctcat
     1981 gcaccaagca cattcaagtt tttgaacttt acccagaacg ttttcacaga tagtattttt
     2041 gaaaaatgtt ccacgttagt taaattggag acacttatct tacaaaagaa tggattaaaa
     2101 gaccttttca aagtaggtct catgacgaag gatatgcctt ctttggaaat actggatgtt
     2161 agctggaatt ctttggaatc tggtagacat aaagaaaact gcacttgggt tgagagtata
     2221 gtggtgttaa atttgtcttc aaatatgctt actgactctg ttttcagatg tttacctccc
     2281 aggatcaagg tacttgatct tcacagcaat aaaataaaga gcgttcctaa acaagtcgta
     2341 aaactggaag ctttgcaaga actcaatgtt gctttcaatt ctttaactga ccttcctgga
     2401 tgtggcagct ttagcagcct ttctgtattg atcattgatc acaattcagt ttcccaccca
     2461 tcggctgatt tcttccagag ctgccagaag atgaggtcaa taaaagcagg ggacaatcca
     2521 ttccaatgta cctgtgagct aagagaattt gtcaaaaata tagaccaagt atcaagtgaa
     2581 gtgttagagg gctggcctga ttcttataag tgtgactacc cagaaagtta tagaggaagc
     2641 ccactaaagg actttcacat gtctgaatta tcctgcaaca taactctgct gatcgtcacc
     2701 atcggtgcca ccatgctggt gttggctgtg actgtgacct ccctctgcat ctacttggat
     2761 ctgccctggt atctcaggat ggtgtgccag tggacccaga ctcggcgcag ggccaggaac
     2821 atacccttag aagaactcca aagaaacctc cagtttcatg cttttatttc atatagtgaa
     2881 catgattctg cctgggtgaa aagtgaattg gtaccttacc tagaaaaaga agatatacag
     2941 atttgtcttc atgagagaaa ctttgtccct ggcaagagca ttgtggaaaa tatcatcaac
     3001 tgcattgaga agagttacaa gtccatcttt gttttgtctc ccaactttgt ccagagtgag
     3061 tggtgccatt acgaactcta ttttgcccat cacaatctct ttcatgaagg atctaataac
     3121 ttaatcctca tcttactgga acccattcca cagaacagca ttcccaacaa gtaccacaag
     3181 ctgaaggctc tcatgacgca gcggacttat ttgcagtggc ccaaggagaa aagcaaacgt
     3241 gggctctttt gggctaacat tagagccgct tttaatatga aattaacact agtcactgaa
     3301 aacaatgatg tgaaatctct cgaggaacaa aaactaatat cagaggaaga tttggaacag
     3361 aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttatgaggg
     3421 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3481 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3541 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3601 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3661 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3721 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3781 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3841 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3901 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3961 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4021 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4081 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4141 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4321 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4441 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4501 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4561 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4621 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4681 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4741 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4801 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4861 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4921 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4981 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5041 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5101 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5161 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5221 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5281 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5341 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5401 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5461 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5521 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5581 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5641 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5701 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6601 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7861 agatctcccg atcccctatg gt
//
