LOCUS       V043227                 8646 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043227
VERSION     V043227
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..3351
                     /label="Tlr2(NM_011905)"
                     /note="Tlr2(NM_011905)"
                     /gene="Tlr2"
     polyA_signal    3405..4027
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        4069..4366
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4420..5214
                     /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    5866..6000
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(6006..6024)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6038..6054)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6062..6078
                     /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(6086..6116)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6131..6152
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6440..7028)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7199..8059)
                     /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(8060..8164)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8191..8619
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgctacgagc tctttggctc
     1021 ttctggatct tggtggccat aacagtcctc ttcagcaaac gctgttctgc tcaggagtct
     1081 ctgtcatgtg atgcttctgg ggtgtgtgat ggccgctcca ggtctttcac ctctattccc
     1141 tccggactca cagcagccat gaaaagcctt gacctgtctt tcaacaagat cacctacatt
     1201 ggccatggtg acctccgagc gtgtgcgaac ctccaggttc tgatgttgaa gtccagcaga
     1261 atcaatacaa tagagggaga cgccttttat tctctgggca gtcttgaaca tttggatttg
     1321 tctgataatc acctatctag tttatcttcc tcctggttcg ggcccctttc ctctttgaaa
     1381 tacttaaact taatgggaaa tccttaccag acactggggg taacatcgct ttttcccaat
     1441 ctcacaaatt tacaaaccct caggatagga aatgtagaga ctttcagtga gataaggaga
     1501 atagattttg ctgggctgac ttctctcaat gaacttgaaa ttaaggcatt aagtctccgg
     1561 aattatcagt cccaaagtct aaagtcgatc cgcgacatcc atcacctgac tcttcactta
     1621 agcgagtctg ctttcctgct ggagattttt gcagatattc tgagttctgt gagatattta
     1681 gaactaagag atactaactt ggccaggttc cagttttcac cactgcccgt agatgaagtc
     1741 agctcaccga tgaagaagct ggcattccga ggctcggttc tcactgatga aagctttaac
     1801 gagctcctga agctgttgcg ttacatcttg gaactgtcgg aggtagagtt cgacgactgt
     1861 accctcaatg ggctcggcga tttcaacccc tcggagtcag acgtagtgag cgagctgggt
     1921 aaagtagaaa cagtcactat ccggaggttg catatccccc agttctattt gttttatgac
     1981 ctgagtactg tctattccct cctggagaag gtgaagcgaa tcacagtaga gaacagcaag
     2041 gtcttcctgg ttccctgctc gttctcccag catttaaaat cattagaatt cttagacctc
     2101 agcgaaaatc tgatggttga agaatatttg aagaactcag cctgtaaggg agcctggcct
     2161 tctctacaaa ccttagtttt gagccagaat catttgagat caatgcaaaa aacaggagag
     2221 attttgctga ctctgaaaaa cctgacctct cttgacatca gcaggaacac ttttcatccg
     2281 atgcccgaca gctgtcagtg gccagaaaag atgcgcttcc tgaatttgtc cagtacaggg
     2341 atccgggtgg taaaaacgtg cattcctcag acgctggagg tgttggatgt tagtaacaac
     2401 aatcttgact cattttcttt gttcttgcct cggctgcaag agctctatat ttccagaaat
     2461 aagctgaaaa cactcccaga tgcttcgttg ttccctgtgt tgctggtcat gaaaatcaga
     2521 gagaatgcag taagtacttt ctctaaagac caacttggtt cttttcccaa actggagact
     2581 ctggaagcag gcgacaacca ctttgtttgc tcctgcgaac tcctatcctt tactatggag
     2641 acgccagctc tggctcaaat cctggttgac tggccagaca gctacctgtg tgactctccg
     2701 cctcgcctgc acggccacag gcttcaggat gcccggccct ccgtcttgga atgtcaccag
     2761 gctgcactgg tgtctggagt ctgctgtgcc cttctcctgt tgatcttgct cgtaggtgcc
     2821 ctgtgccacc atttccacgg actgtggtac ctgagaatga tgtgggcgtg gctccaggcc
     2881 aagaggaagc ccaagaaagc tccctgcagg gacgtttgct atgatgcctt tgtttcctac
     2941 agtgagcagg attcccattg ggtggagaac ctcatggtcc agcagctgga gaactctgac
     3001 ccgcccttta agctgtgtct ccacaagcgg gacttcgttc cgggcaaatg gatcattgac
     3061 aacatcatcg attccatcga aaagagccac aaaactgtgt tcgtgctttc tgagaacttc
     3121 gtacggagcg agtggtgcaa gtacgaactg gacttctccc acttcaggct ctttgacgag
     3181 aacaacgacg cggccatcct tgttttgctg gagcccattg agaggaaagc cattccccag
     3241 cgcttctgca aactgcgcaa gataatgaac accaagacct acctggagtg gcccttggat
     3301 gaaggccagc aggaagtgtt ttgggtaaat ctgagaactg caataaagtc ctaggaattc
     3361 atcgatagat ctgatatcgg taccagtcga ctctagagga tcccgggtgg catccctgtg
     3421 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     3481 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3541 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3601 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     3661 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3721 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     3781 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     3841 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat
     3901 accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac
     3961 atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg
     4021 ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4081 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4141 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4201 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4321 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggagga attgatcagc
     4381 ttgggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4441 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4501 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4561 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4621 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4681 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4741 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4801 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4861 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4921 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4981 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5041 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5101 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5161 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5221 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5281 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5341 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccggg ctcgatcccc
     5401 tcgcgagttg gttcagctgc tgcctgaggc tggacgacct cgcggagttc taccggcagt
     5461 gcaaatccgt cggcatccag gaaaccagca gcggctatcc gcgcatccat gcccccgaac
     5521 tgcaggagtg gggaggcacg atggccgctt tggtcgaccc ggacgggacg ctcctgcgcc
     5581 tgatacagaa cgaattgctt gcaggcatct catgagtgtg tcttcccgtt ttccgcctga
     5641 ggtcactgcg tggatggagc gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca
     5701 ctcgccaagc tggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccg
     5761 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     5821 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     6001 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     6061 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6121 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6181 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6241 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6301 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6361 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6421 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6481 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6541 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6601 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6661 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6721 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6781 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6841 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6901 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6961 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7021 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7081 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7141 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7201 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7261 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7321 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7381 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7441 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7501 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7561 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7621 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7681 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7741 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7801 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7861 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7921 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7981 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8041 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8101 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8161 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     8221 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     8281 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8341 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8401 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8461 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8521 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     8581 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     8641 caattt
//
