LOCUS       V042239                 8569 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042239
VERSION     V042239
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             964..4059
                     /label="Tlr9(NM_031178)"
                     /note="Tlr9(NM_031178)"
                     /gene="Tlr9"
     CDS             4066..4131
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4169..4393
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4439..4867
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4881..5210
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5277..6071
                     /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    6245..6366
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6415..6431)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6439..6455
                     /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(6463..6493)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6508..6529
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6817..7402)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7573..8433)
                     /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(8434..8538)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggttc tccgtcgaag gactctgcac cccttgtccc tcctggtaca ggctgcagtg
     1021 ctggctgaga ctctggccct gggtaccctg cctgccttcc taccctgtga gctgaagcct
     1081 catggcctgg tggactgcaa ttggctgttc ctgaagtctg taccccgttt ctctgcggca
     1141 gcatcctgct ccaacatcac ccgcctctcc ttgatctcca accgtatcca ccacctgcac
     1201 aactctgact tcgtccacct gtccaacctg cggcagctga acctcaagtg gaactgtcca
     1261 cccactggcc ttagccccct gcacttctct tgccacatga ccattgagcc cagaaccttc
     1321 ctggctatgc gtacactgga ggagctgaac ttgagctata atggtatcac cactgtgccc
     1381 cgactgccca gctccctggt gaatctgagc ctgagccaca ccaacatcct ggttctagat
     1441 gctaacagcc tcgccggcct atacagcctg cgcgttctct tcatggacgg gaactgctac
     1501 tacaagaacc cctgcacagg agcggtgaag gtgaccccag gcgccctcct gggcctgagc
     1561 aatctcaccc atctgtctct gaagtataac aacctcacaa aggtgccccg ccaactgccc
     1621 cccagcctgg agtacctcct ggtgtcctat aacctcattg tcaagctggg gcctgaagac
     1681 ctggccaatc tgacctccct tcgagtactt gatgtgggtg ggaattgccg tcgctgtgac
     1741 catgccccca atccctgtat agaatgtggc caaaagtccc tccacctgca ccctgagacc
     1801 ttccatcacc tgagccatct ggaaggcctg gtgctgaagg acagctctct ccatacactg
     1861 aactcttcct ggttccaagg tctggtcaac ctctcggtgc tggacctaag cgagaacttt
     1921 ctctatgaaa gcatcaccca caccaatgcc tttcagaacc taacccgcct gcgcaagctc
     1981 aacctgtcct tcaattaccg caagaaggta tcctttgccc gcctccacct ggcaagttcc
     2041 tttaagaacc tggtgtcact gcaggagctg aacatgaacg gcatcttctt ccgcttgctc
     2101 aacaagtaca cgctcagatg gctggccgat ctgcccaaac tccacactct gcatcttcaa
     2161 atgaacttca tcaaccaggc acagctcagc atctttggta ccttccgagc ccttcgcttt
     2221 gtggacttgt cagacaatcg catcagtggg ccttcaacgc tgtcagaagc cacccctgaa
     2281 gaggcagatg atgcagagca ggaggagctg ttgtctgcgg atcctcaccc agctccgctg
     2341 agcacccctg cttctaagaa cttcatggac aggtgtaaga acttcaagtt caccatggac
     2401 ctgtctcgga acaacctggt gactatcaag ccagagatgt ttgtcaatct ctcacgcctc
     2461 cagtgtctta gcctgagcca caactccatt gcacaggctg tcaatggctc tcagttcctg
     2521 ccgctgacta atctgcaggt gctggacctg tcccataaca aactggactt gtaccactgg
     2581 aaatcgttca gtgagctacc acagttgcag gccctggacc tgagctacaa cagccagccc
     2641 tttagcatga agggtatagg ccacaatttc agttttgtga cccatctgtc catgctacag
     2701 agccttagcc tggcacacaa tgacattcat acccgtgtgt cctcacatct caacagcaac
     2761 tcagtgaggt ttcttgactt cagcggcaac ggtatgggcc gcatgtggga tgaggggggc
     2821 ctttatctcc atttcttcca aggcctgagt ggcctgctga agctggacct gtctcaaaat
     2881 aacctgcata tcctccggcc ccagaacctt gacaacctcc ccaagagcct gaagctgctg
     2941 agcctccgag acaactacct atctttcttt aactggacca gtctgtcctt cctacccaac
     3001 ctggaagtcc tagacctggc aggcaaccag ctaaaggccc tgaccaatgg caccctgcct
     3061 aatggcaccc tcctccagaa actcgatgtc agtagcaaca gtatcgtctc tgtggtccca
     3121 gccttcttcg ctctggcggt cgagctgaaa gaggtcaacc tcagccacaa cattctcaag
     3181 acggtggatc gctcctggtt tgggcccatt gtgatgaacc tgacagttct agacgtgaga
     3241 agcaaccctc tgcactgtgc ctgtggggca gccttcgtag acttactgtt ggaggtgcag
     3301 accaaggtgc ctggcctggc taatggtgtg aagtgtggca gccccggcca gctgcagggc
     3361 cgtagcatct tcgcgcagga cctgcggctg tgcctggatg aggtcctctc ttgggactgc
     3421 tttggccttt cactcttggc tgtggccgtg ggcatggtgg tgcctatact gcaccatctc
     3481 tgcggctggg acgtctggta ctgttttcat ctgtgcctgg catggctacc tttgctagcc
     3541 cgcagccgac gcagcgccca aactctccct tatgatgcct tcgtggtgtt cgataaggca
     3601 cagagcgcag ttgccgactg ggtgtataac gagctgcggg tgcggctgga ggagcggcgc
     3661 ggccgccgag ccctacgctt gtgtctggag gaccgagatt ggctgcctgg ccagacgctc
     3721 ttcgagaacc tctgggcttc catctatggg agccgcaaga ctctatttgt gctggcccac
     3781 acggaccgcg tcagtggcct cctgcgcacc agcttcctgc tggctcagca gcgcctgttg
     3841 gaagaccgca aggacgtggt ggtgttggtg atcctgcgtc cggatgccca ccgctcccgc
     3901 tatgtgcgac tgcgccagcg tctctgccgc cagagtgtgc tcttctggcc ccagcagccc
     3961 aacgggcagg ggggcttctg ggcccagctg agtacagccc tgactaggga caaccgccac
     4021 ttctataacc agaacttctg ccggggacct acagcagaac tcgaggacta caaagaccat
     4081 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg
     4141 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4201 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4261 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4321 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4381 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4441 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4501 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4561 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4621 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4681 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4741 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4801 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4861 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4981 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5041 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5101 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5161 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5221 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5281 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5341 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5401 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5461 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5521 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5581 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5641 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5701 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5761 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5821 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5881 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5941 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6001 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6061 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6121 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6181 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6241 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6361 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6421 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6481 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6541 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7321 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     7381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7561 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7741 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8101 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8401 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
