LOCUS       V027800                 8718 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027800
VERSION     V027800
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3398
                     /label="Nlrp6(NM_134375)"
                     /note="Nlrp6(NM_134375)"
                     /gene="Nlrp6"
     CDS             3405..3470
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3494..4082
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4148..4196
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4398..4826
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4840..5169
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5236..6030
                     /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    6206..6327
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6376..6392)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6400..6416
                     /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(6424..6454)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6469..6490
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6778..7366)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7537..8397)
                     /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(8398..8502)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga tgccgctgga gcctcctgct
      781 ccagctcgga ggcggtggca agggaactgc ttctggccgc actgcaggat ctgagccagg
      841 agcagctgaa gcgttttcgc cacaagctcc gggatgcgcc cctggatggc cgcagcatcc
      901 cgtgggggcg actggagcac tcagacgctg tggacctcac agacaagctg attgagttct
      961 atgcgccaga gccagccgta gatgtgaccc gaaagatcct gaagaaggcg gatattcgag
     1021 atgtgtcgtt gcgactcaag gagcagcagc tgcagaggct cggctccagc tcggcgcttc
     1081 tcaccgtgtc ggagtacaag aagaagtacc gcgagcacgt gctacgacag catgctaagg
     1141 tgaaggagag gaatgcccgc tccgtgaaga tcaacaagcg cttcaccaag ctgctcattg
     1201 cgccggggag cggcgccggg gaggacgagc tgctgggaac gtcgggggag cccgagcccg
     1261 agcgcgcaag gcgctcggac acacacactt tcaatcggct cttccgagga aatgatgacg
     1321 agggcccgcg gccgctgacc gtggtgctgc agggcccggc aggcatcggc aaaaccatgg
     1381 ccgccaagaa aatcctctac gactgggcag gaggcaagct ataccacagc caagtggact
     1441 ttgccttctt catgccgtgc ggcgagctgc tggagcggcc aggtacgcgc agcttggcgg
     1501 acctgatact ggaacagtgc ccggaccgca ccgcgccggt gcggcggata ctagcgcaac
     1561 cgcatcgtct actgttcatc ctggacggtg cagacgaatt gcctacttta gcagccccag
     1621 aagccactcc ctgcagagac cctttcgagg ctacgagtgg cttgagagtg ctgagcgggc
     1681 tgctgagcca ggagctgtta ccatcggctc gcttgctggt gacttcacgc aacgccaccc
     1741 tcgggaggct gcagggcaga ctttgctcac cacagtgcgc agaagtgcgc ggcttctcag
     1801 acaaagacaa gaaaaagtat ttcttcaagt ttttccggga cgagaggaag gcagagcgcg
     1861 cttaccgctt cgtgaaagag aatgagacgc tgtacgcact gtgctttgtg ccgtttgtgt
     1921 gctggatcgt gtgcaccgtg ctgctgcagc aaatggagct gggccgggat ctgtctcgta
     1981 cctccaagac cactacatct gtgtacctgc tcttcatcac cagcatgctg aagtctgcag
     2041 gtaccaatgg accccgggtt cagggagagc tgcgaatgct gtgccgcctg gcccgggagg
     2101 gcatcctgaa gcatcaagca cagttctcag aaaaggacct ggagagattg aagcttcagg
     2161 gttcccaagt tcagacaatg tttctcagca agaaggagct gccaggagtg ctagaaactg
     2221 tggtcaccta ccagttcatt gaccagagct tccaggagtt cttggctgca ttgtcatacc
     2281 tactagacgc tgagggagcc ccagggaact ccgcaggaag tgtgcagatg ctcctgaact
     2341 ctgacgcggg gctgcgtggt catctggcac tcaccactag attcctcttt ggactgctaa
     2401 gtacagagag gattcgtgac attggaaacc attttggctg tgtggtgcca gggcgtgtga
     2461 aacaggacac cttgcggtgg gtacaaggac aaagccaacc caaggtggcg acagtaggag
     2521 cagaaaagaa ggatgagctg aaggacgagg aagcagagga ggaggaggag gaggaggaag
     2581 aagaggagga ggaactcaac tttggactgg agctgttgta ctgcctgtat gagacacaag
     2641 aggatgattt tgttcgccag gctctcagca gccttccaga gatggtactg gagcgagtta
     2701 ggctgacccg catggacctt gaggttctga gctactgcgt gcagtgctgc ccggacggcc
     2761 aggctctgag actggtgagc tgtggactgg tggcggcaaa ggagaagaag aagaagaaga
     2821 agagcttcat gaaccggctg aagggttctc aaagcaccgg gaaacaaccc ccagcctcct
     2881 tgctgcgtcc actctgtgag gcaatgatta cccagcaatg tggtctgagt attctgacct
     2941 tgtcacactg caaactccct gatgcagttt gtcgagacct ttctgaggct ctgaaggtag
     3001 ctccttccct aagggagctg ggcctcctcc agaaccgact cactgaggca ggcctgcgtt
     3061 tactaagcca aggcctggct tggcccaaat gcaaggtgca gacactcagg atacagatgc
     3121 ctggcctcca agaggtgatc cactacctgg tcattgtgct ccagcagagc ccagtcctaa
     3181 ccactctgga cctcagtggc tgtcagctgc ctggcactgt ggtggaacct ctgtgttcag
     3241 ccctgaagca ccctaaatgt ggcctaaaga ccctcagtct gacttctgtg gagctgactg
     3301 agaatccact gagagagctt caagctgtga agacattaaa gccagatctg gccatcatac
     3361 attcaaaatt gggcacacat cctcagcctc tgaagggagg atccgactac aaagaccatg
     3421 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     3481 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3541 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3601 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3661 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3721 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3781 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3841 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3901 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3961 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4021 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4081 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4141 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4201 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4261 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4321 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4381 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4441 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4501 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     4561 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4621 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4681 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4741 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4801 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4861 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4921 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4981 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5041 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5101 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5161 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5221 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5281 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5341 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5401 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5461 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5521 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5581 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5641 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5701 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5761 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5821 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5881 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5941 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6001 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6061 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6121 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6181 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6241 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6301 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6361 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6421 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6481 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6541 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6601 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6661 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6721 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6781 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6841 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6901 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6961 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7021 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7081 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7141 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7201 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7261 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7321 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7381 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7441 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7501 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7561 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7621 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7681 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7741 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7801 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7861 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7921 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7981 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8041 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8101 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8161 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8221 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8281 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8341 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8401 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8461 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8521 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     8581 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8641 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8701 atgaagaatc tgcttagg
//
