LOCUS       V018713                 8720 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018713
VERSION     V018713
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..3400
                     /label="TMC2(NM_080751)"
                     /note="TMC2(NM_080751)"
                     /gene="TMC2"
     CDS             3407..3472
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3496..4084
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4150..4198
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4400..4828
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4842..5171
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5238..6032
                     /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    6208..6329
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6378..6394)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6402..6418
                     /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(6426..6456)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6471..6492
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6780..7368)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7539..8399)
                     /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(8400..8504)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgagcca ccaggtaaag ggcctgaaag aggaagcacg
      721 aggcggagtg aaagggcggg tgaagagcgg ctctccacac acaggtgaca ggctgggaag
      781 gagatcctca agcaagcggg ctctcaaagc cgaggggacc ccaggcaggc gcggagctca
      841 gcgaagccag aaggagcgcg ccgggggcag cccaagcccg gggtctcccc ggaggaagca
      901 aacagggcgc aggagacaca gagaagagct gggggagcag gagcggggcg aggcagagag
      961 gacctgcgag ggcaggagaa agcgcgacga gagggcctcc ttccaggagc ggacagcagc
     1021 cccaaagagg gaaaaggaga ttccgaggag ggaggagaag tcgaagcggc agaagaaacc
     1081 caggtcatcc tccttggcct ccagtgcctc tggtggggag tccctgtccg aggaggaact
     1141 ggcccagatc ctggagcagg tggaagaaaa aaagaagctc attgccacca tgcggagcaa
     1201 gccctggccc atggcgaaga agctgacaga gctcagggag gcccaggaat ttgtggagaa
     1261 gtatgaaggt gccttgggaa aggggaaagg caagcaacta tatgcctaca agatgctgat
     1321 ggccaagaaa tgggtcaaat ttaagagaga ctttgataat ttcaagactc aatgtatccc
     1381 ctgggaaatg aagatcaagg acattgaaag tcactttggt tcttcagtgg catcgtattt
     1441 catctttctc cgatggatgt atggagttaa ccttgtcctt tttggcttaa tatttggtct
     1501 agtcataatc ccagaggtac tgatgggcat gccctatggg agtattccca gaaagacagt
     1561 gcctcgggct gaggaagaaa aggccatgga tttttctgtc ctttgggatt ttgagggcta
     1621 tatcaagtac tctgcactct tctatggcta ctacaacaac cagaggacca tcgggtggct
     1681 gaggtaccgg ctgcctatgg cttactttat ggtgggggtc agcgtgttcg gctacagcct
     1741 gattattgtc attcgatcga tggccagcaa tacccaagga agcacaggcg aaggggagag
     1801 tgacaacttc acattcagct tcaagatgtt caccagctgg gactacctga tcgggaattc
     1861 agagacagct gataacaaat atgcatccat caccaccagc ttcaaggaat caatagtgga
     1921 tgaacaagag agtaacaaag aagaaaatat ccatctgaca agatttcttc gtgtcctggc
     1981 caactttctc atcatctgct gtttgtgtgg aagtgggtac ctcatttact ttgtggttaa
     2041 gcgatctcag caattctcca aaatgcagaa tgtcagctgg tatgaaagga atgaggtaga
     2101 gatcgtgatg tccctgcttg gaatgttttg tccccctctg tttgaaacca tcgctgccct
     2161 ggagaattac cacccacgca ctggactgaa gtggcagctg ggacgcatct ttgcactctt
     2221 cctggggaac ctctacacat ttctcttggc cctgatggat gacgtccacc tcaagcttgc
     2281 taatgaagag acaataaaga acatcactca ctggactctg tttaactatt acaactcttc
     2341 tggttggaac gagagtgtcc cccgaccacc cctgcaccct gcagatgtgc cccggggttc
     2401 ttgctgggag acagctgtgg gcattgaatt catgaggctg acggtgtctg acatgctggt
     2461 aacgtacatc accatcctgc tgggggactt cctacgggct tgttttgtgc ggttcatgaa
     2521 ctactgctgg tgctgggact tggaggctgg atttccttca tatgctgagt ttgatattag
     2581 tggaaatgtg ctgggtttga tcttcaacca aggaatgatc tggatgggct ccttctatgc
     2641 tccaggcctg gtgggcatta atgtgctgcg cctgctgacc tccatgtact tccagtgctg
     2701 ggcggtgatg agcagcaacg taccccatga acgcgtgttc aaagcctccc gatccaacaa
     2761 cttctacatg ggcctcctgc tgctggtgct cttcctcagc ctcctgccgg tggcctacac
     2821 catcatgtcc ctcccaccct cctttgactg cgggccgttc agtgggaaaa acagaatgta
     2881 cgatgtcctc caagagacca ttgaaaacga tttcccaacc ttcctgggca agatctttgc
     2941 tttcctcgcc aatccaggcc tgatcatccc agccatcctg ctgatgttct tggccattta
     3001 ctacctgaac tcagtttcca aaagcctttc ccgagctaat gcccagctga ggaagaaaat
     3061 ccaagtgctc cgtgaagttg agaagagtca caaatctgta aaaggcaaag ccacagccag
     3121 agattcagag gacacaccta aaagcagctc caaaaatgcc acccagctcc aactcaccaa
     3181 ggaagagacc actcctccct ctgccagcca aagccaggcc atggacaaga aggcgcaggg
     3241 ccctgggacc tccaattctg ccagcaggac cacactgcct gcctctggac accttcctat
     3301 atctcggccc cctggaatcg gaccagattc tggccacgcc ccatctcaga ctcatccgtg
     3361 gaggtcagcc tctggaaaga gtgctcagag acctccccac ctcgaggact acaaagacca
     3421 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     3481 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3541 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3601 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3661 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3721 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3781 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3841 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     3901 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3961 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4021 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4081 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4141 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     4201 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4261 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4321 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     4381 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4441 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4501 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     4561 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4621 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4681 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4741 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     4801 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4921 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4981 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5041 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5101 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5161 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     5221 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5281 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5341 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5401 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5461 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5521 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5581 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5641 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5701 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5761 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5821 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5881 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5941 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6001 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     6061 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6121 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6181 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6301 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6361 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6421 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6481 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6541 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6601 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6661 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6721 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6781 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6841 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6901 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6961 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7021 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7081 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7141 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7201 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7261 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7321 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7381 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7441 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7501 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7561 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7621 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7681 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7741 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7801 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7861 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7921 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7981 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8041 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8101 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8161 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8221 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8281 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8341 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8401 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8461 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8521 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     8581 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     8641 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     8701 gcatgaagaa tctgcttagg
//
