LOCUS       V018562                 7961 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018562
VERSION     V018562
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..3501
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        complement(3514..3532)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    3558..3782
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3828..4256
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4270..4599
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4666..5460
                     /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    5634..5755
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5804..5820)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5828..5844
                     /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(5852..5882)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5897..5918
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6206..6794)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6965..7825)
                     /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(7826..7930)
                     /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 tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3481 tacaaggatg acgatgacaa gtgagggccc tattctatag tgtcacctaa atgctagagc
     3541 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3601 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3661 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3721 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3781 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3901 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4021 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4141 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4201 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4261 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4441 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4501 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4561 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4621 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4681 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4741 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4801 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4861 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4921 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4981 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5041 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5101 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5161 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5221 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5281 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5341 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5401 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5461 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5521 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5581 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5641 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5701 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5761 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6001 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6061 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
