LOCUS       V029052                 7913 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029052
VERSION     V029052
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             913..3429
                     /label="TLR4(NM_138554)"
                     /note="TLR4(NM_138554)"
                     /gene="TLR4"
     CDS             3436..3453
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        complement(3466..3484)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    3510..3734
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3780..4208
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4222..4551
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4618..5412
                     /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    5586..5707
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5756..5772)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5780..5796
                     /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(5804..5834)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5849..5870
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6158..6746)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6917..7777)
                     /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(7778..7882)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccatgatgtc tgcctcgcgc ctggctggga ctctgatccc agccatggcc
      961 ttcctctcct gcgtgagacc agaaagctgg gagccctgcg tggaggtggt tcctaatatt
     1021 acttatcaat gcatggagct gaatttctac aaaatccccg acaacctccc cttctcaacc
     1081 aagaacctgg acctgagctt taatcccctg aggcatttag gcagctatag cttcttcagt
     1141 ttcccagaac tgcaggtgct ggatttatcc aggtgtgaaa tccagacaat tgaagatggg
     1201 gcatatcaga gcctaagcca cctctctacc ttaatattga caggaaaccc catccagagt
     1261 ttagccctgg gagccttttc tggactatca agtttacaga agctggtggc tgtggagaca
     1321 aatctagcat ctctagagaa cttccccatt ggacatctca aaactttgaa agaacttaat
     1381 gtggctcaca atcttatcca atctttcaaa ttacctgagt atttttctaa tctgaccaat
     1441 ctagagcact tggacctttc cagcaacaag attcaaagta tttattgcac agacttgcgg
     1501 gttctacatc aaatgcccct actcaatctc tctttagacc tgtccctgaa ccctatgaac
     1561 tttatccaac caggtgcatt taaagaaatt aggcttcata agctgacttt aagaaataat
     1621 tttgatagtt taaatgtaat gaaaacttgt attcaaggtc tggctggttt agaagtccat
     1681 cgtttggttc tgggagaatt tagaaatgaa ggaaacttgg aaaagtttga caaatctgct
     1741 ctagagggcc tgtgcaattt gaccattgaa gaattccgat tagcatactt agactactac
     1801 ctcgatgata ttattgactt atttaattgt ttgacaaatg tttcttcatt ttccctggtg
     1861 agtgtgacta ttgaaagggt aaaagacttt tcttataatt tcggatggca acatttagaa
     1921 ttagttaact gtaaatttgg acagtttccc acattgaaac tcaaatctct caaaaggctt
     1981 actttcactt ccaacaaagg tgggaatgct ttttcagaag ttgatctacc aagccttgag
     2041 tttctagatc tcagtagaaa tggcttgagt ttcaaaggtt gctgttctca aagtgatttt
     2101 gggacaacca gcctaaagta tttagatctg agcttcaatg gtgttattac catgagttca
     2161 aacttcttgg gcttagaaca actagaacat ctggatttcc agcattccaa tttgaaacaa
     2221 atgagtgagt tttcagtatt cctatcactc agaaacctca tttaccttga catttctcat
     2281 actcacacca gagttgcttt caatggcatc ttcaatggct tgtccagtct cgaagtcttg
     2341 aaaatggctg gcaattcttt ccaggaaaac ttccttccag atatcttcac agagctgaga
     2401 aacttgacct tcctggacct ctctcagtgt caactggagc agttgtctcc aacagcattt
     2461 aactcactct ccagtcttca ggtactaaat atgagccaca acaacttctt ttcattggat
     2521 acgtttcctt ataagtgtct gaactccctc caggttcttg attacagtct caatcacata
     2581 atgacttcca aaaaacagga actacagcat tttccaagta gtctagcttt cttaaatctt
     2641 actcagaatg actttgcttg tacttgtgaa caccagagtt tcctgcaatg gatcaaggac
     2701 cagaggcagc tcttggtgga agttgaacga atggaatgtg caacaccttc agataagcag
     2761 ggcatgcctg tgctgagttt gaatatcacc tgtcagatga ataagaccat cattggtgtg
     2821 tcggtcctca gtgtgcttgt agtatctgtt gtagcagttc tggtctataa gttctatttt
     2881 cacctgatgc ttcttgctgg ctgcataaag tatggtagag gtgaaaacat ctatgatgcc
     2941 tttgttatct actcaagcca ggatgaggac tgggtaagga atgagctagt aaagaattta
     3001 gaagaagggg tgcctccatt tcagctctgc cttcactaca gagactttat tcccggtgtg
     3061 gccattgctg ccaacatcat ccatgaaggt ttccataaaa gccgaaaggt gattgttgtg
     3121 gtgtcccagc acttcatcca gagccgctgg tgtatctttg aatatgagat tgctcagacc
     3181 tggcagtttc tgagcagtcg tgctggtatc atcttcattg tcctgcagaa ggtggagaag
     3241 accctgctca ggcagcaggt ggagctgtac cgccttctca gcaggaacac ttacctggag
     3301 tgggaggaca gtgtcctggg gcggcacatc ttctggagac gactcagaaa agccctgctg
     3361 gatggtaaat catggaatcc agaaggaaca gtgggtacag gatgcaattg gcaggaagca
     3421 acatctatcc tcgagcatca tcatcatcat cattgagggc cctattctat agtgtcacct
     3481 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3541 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3601 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3661 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3721 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3841 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3961 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4081 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4141 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4201 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4261 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4321 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4381 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4441 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4501 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4561 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4621 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4681 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4741 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4801 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4861 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4921 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4981 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5041 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5101 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5161 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5221 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5281 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5341 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5401 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5461 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5521 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5581 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5641 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5701 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5761 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5821 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5881 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
