LOCUS       V036087                 9379 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036087
VERSION     V036087
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             706..4038
                     /label="Nlrp5(NM_011860)"
                     /note="Nlrp5(NM_011860)"
                     /gene="Nlrp5"
     CDS             4045..4071
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4075..4101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4105..4131
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4155..4743
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4809..4857
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5059..5487
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5501..5830
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5897..6691
                     /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    6867..6988
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7037..7053)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7061..7077
                     /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(7085..7115)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7130..7151
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7439..8027)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8198..9058)
                     /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(9059..9163)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggg tcctccagaa
      721 aaagaaagta aagcaatctt gaaagcacgt ggattggaag aggaacagaa gtcagaaaga
      781 aaaatgactt ctccagaaaa cgacagtaaa tcaatccaga aagaccaagg accagagcag
      841 gagcagacat cagaaagcac aatgggtcct ccagaaaaag acagtaaagc aatcttgaaa
      901 gcacgtggat tggaagagga acagaagtca gaaagcacaa tgtctccttc agaaaatgtc
      961 agtagagcaa tcctgaaaga cagtggatca gaagaagtgg aacaggcgtc agaaagaaaa
     1021 atgacttctc cagaaaacga cagtaaatca atccagaaag accaaggacc agagcaggag
     1081 cagacatcag aaaccttaca atctaaggaa gaagatgaag tgacagaggc agataaagat
     1141 aatggaggtg acttacaaga ctacaaggcc catgtgattg ctaagttcga cacaagtgtg
     1201 gatctacact atgacagccc agagatgaaa ttattgtctg atgcttttaa accataccag
     1261 aaaaccttcc agcctcacac cattatccta catggaagac caggagttgg gaagtcagct
     1321 ttggccagaa gtattgttct tggctgggca cagggtaaac tcttccaaaa aatgtccttt
     1381 gtcatcttct tctctgttag agaaataaag tggacagaga agagcagttt ggcacagctg
     1441 attgctaagg agtgtccaga ctcctgggat ctagtgacaa agatcatgtc ccaaccagaa
     1501 agactcttgt ttgtcataga tggcttggat gatatggact ctgtcctcca acatgatgat
     1561 atgacactat ccagagactg gaaggatgaa cagcccatat acatcctgat gtacagcctc
     1621 ctgaggaagg ctctcctacc tcagtccttt ctcatcatta ccaccagaaa cacaggctta
     1681 gaaaaactca agtcaatggt tgtgtccccc ctctatatac tggttgaagg actgtctgca
     1741 tcaaggagat ctcagctggt cctcgagaac atctccaatg agtctgatag aatacaagtc
     1801 ttccattctc tgatagaaaa tcaccagctg tttgaccaat gccaggcccc ctctgtgtgc
     1861 tccctggtct gtgaggctct acagctacag aagaaactgg gaaagagatg caccctaccc
     1921 tgccagactc tcaccggttt gtatgccacg ttggtgtttc accagctcac cttgaaaagg
     1981 ccttcccaga gcgctctcag tcaggaagaa cagattactc tagtgggttt gtgcatgatg
     2041 gcagctgaag gagtgtggac catgaggtcg gtgttctatg atgatgacct gaagaactat
     2101 agcctaaagg agtctgagat cttggccctc tttcacatga acatccttct ccaggttggc
     2161 cacaacagtg agcagtgtta tgttttctcc cacctcagcc tgcaggattt ctttgctgcc
     2221 ttatattatg ttttagaagg gctggaggaa tggaatcagc atttttgctt cattgaaaac
     2281 caaaggagca tcatggaggt gaagagaact gacgacactc gcctcctcgg gatgaagcgt
     2341 ttcttatttg gcctcatgaa caaggatatc ttgaagactc tggaggttct gtttgaatat
     2401 cccgtgattc caactgttga gcagaagctc caacactggg tctctctgat agctcagcag
     2461 gtcaatggca ccagcccaat ggacaccctg gatgccttct attgtctatt tgagtctcag
     2521 gatgaagagt ttgttggcgg ggctctcaaa cgcttccaag aagtgtggct gctgattaac
     2581 cagaagatgg acttgaaggt ctcttcctac tgtctcaagc actgtcagaa cttgaaggca
     2641 atccgggtgg atatcagaga cctcctctcg gtagataata ctctcgagct gtgccctgtt
     2701 gttactgtcc aggagacaca atgtaagccc ctcctcatgg agtggtgggg aaacttctgc
     2761 tctgtgcttg gcagcctccg gaacttgaag gagctggact tgggcgacag catcctgagt
     2821 caacgggcca tgaagatact gtgcctcgag ctgcggaatc agtcctgcag aatacagaag
     2881 ctgacgttta agagtgcaga ggtagtgtct ggcctgaaac atctctggaa gctccttttt
     2941 agcaatcaaa acttaaagta cctcaatcta gggaacactc ccatgaagga tgatgacatg
     3001 aagttagcct gcgaagcgct gaaacatcca aagtgctccg tggagactct gaggttggat
     3061 tcctgtgagt taaccatcat tggttatgag atgatctcca cgcttcttat ttcaaccacc
     3121 aggctaaagt gtctcagcct ggccaaaaat agagtgggag taaaaagcat gatatccctt
     3181 gggaatgcct tgagtagctc aatgtgtcta ctgcaaaagt tgatactgga caactgtggc
     3241 ctcacacctg ccagctgcca ccttctggtc tcagcccttt tcagcaacca gaacttgaca
     3301 cacctgtgcc tgtcaaacaa cagcctgggg actgaaggag tgcaacagct gtgtcagttc
     3361 ctgaggaatc cagaatgtgc tctccagcgg ctgatactga atcactgcaa cattgtagat
     3421 gatgcttatg gcttcctggc aatgagactt gcaaacaaca caaagctgac ccacctgagc
     3481 ctgaccatga accccgtagg ggatggtgca atgaagctac tgtgtgaagc tttaaaggaa
     3541 cctacttgtt accttcaaga actggaacta gtggactgcc aactcacaca gaactgctgc
     3601 gaggacctgg cctgtatgat cacaacaacc aagcacttaa aaagtttgga tcttggtaac
     3661 aacgccctgg gtgacaaagg agtcataacc ctgtgtgagg gactgaagca aagtagcagc
     3721 tccctgagga gacttgggtt gggggcatgt aagttgactt ccaattgctg tgaggcattg
     3781 tcattggcca tctcttgcaa ccctcacctg aacagcctaa acctggtgaa gaatgacttc
     3841 agtacatcgg ggatgttgaa gctgtgctct gcgttccaat gccctgtctc taacctgggg
     3901 ataattggcc tgtggaagca ggagtactat gcccgagtga gaagacagct ggaggaagtt
     3961 gagtttgtca agccccacgt ggtgattgat ggtgattggt atgctagtga tgaagatgac
     4021 cgaaactggt ggaaaaacct cgagtaccca tacgacgtac ctgattatgc aacttatcct
     4081 tatgatgtac ctgattatgc tagctatcct tatgatgtcc cagactacgc atagtgaggg
     4141 cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4201 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4261 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     4321 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     4381 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     4441 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     4501 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     4561 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     4621 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4681 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4741 cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4801 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     4861 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4921 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4981 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg
     5041 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5101 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5161 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     5221 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5281 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5341 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5401 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     5461 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     5521 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5581 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5641 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5701 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5761 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5821 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5881 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5941 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     6001 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     6061 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     6121 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     6181 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     6241 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6301 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6361 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6421 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6481 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6541 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6601 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6661 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     6721 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6781 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6841 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6961 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     7021 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7081 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7141 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7621 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7861 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9181 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac
     9241 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     9301 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     9361 catgaagaat ctgcttagg
//
