LOCUS       V023166                10206 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023166
VERSION     V023166
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..4880
                     /label="NLRP1(NM_001033053)"
                     /note="NLRP1(NM_001033053)"
                     /gene="NLRP1"
     CDS             4893..4958
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4982..5570
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5636..5684
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5886..6314
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6328..6657
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6724..7518
                     /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    7694..7815
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7864..7880)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7888..7904
                     /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(7912..7942)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7957..7978
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8266..8854)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9025..9885)
                     /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(9886..9990)
                     /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 ctcggatccg ccaccatggc tggcggagcc tggggccgcc
      781 tggcctgtta cttggagttc ctgaagaagg aggagctgaa ggagttccag cttctgctcg
      841 ccaataaagc gcactccagg agctcttcgg gtgagacacc cgctcagcca gagaagacga
      901 gtggcatgga ggtggcctcg tacctggtgg ctcagtatgg ggagcagcgg gcctgggacc
      961 tagccctcca tacctgggag cagatggggc tgaggtcact gtgcgcccaa gcccaggaag
     1021 gggcaggcca ctctccctca ttcccctaca gcccaagtga accccacctg gggtctccca
     1081 gccaacccac ctccaccgca gtgctaatgc cctggatcca tgaattgccg gcggggtgca
     1141 cccagggctc agagagaagg gttttgagac agctgcctga cacatctgga cgccgctgga
     1201 gagaaatctc tgcctcactc ctctaccaag ctcttccaag ctccccagac catgagtctc
     1261 caagccagga gtcacccaac gcccccacat ccacagcagt gctggggagc tggggatccc
     1321 cacctcagcc cagcctagca cccagagagc aggaggctcc tgggacccaa tggcctctgg
     1381 atgaaacgtc aggaatttac tacacagaaa tcagagaaag agagagagag aaatcagaga
     1441 aaggcaggcc cccatgggca gcggtggtag gaacgccccc acaggcgcac accagcctac
     1501 agccccacca ccacccatgg gagccttctg tgagagagag cctctgttcc acatggccct
     1561 ggaaaaatga ggattttaac caaaaattca cacagctgct acttctacaa agacctcacc
     1621 ccagaagcca agatcccctg gtcaagagaa gctggcctga ttatgtggag gagaatcgag
     1681 gacatttaat tgagatcaga gacttatttg gcccaggcct ggatacccaa gaacctcgca
     1741 tagtcatact gcagggggct gctggaattg ggaagtcaac actggccagg caggtgaagg
     1801 aagcctgggg gagaggccag ctgtatgggg accgcttcca gcatgtcttc tacttcagct
     1861 gcagagagct ggcccagtcc aaggtggtga gtctcgctga gctcatcgga aaagatggga
     1921 cagccactcc ggctcccatt agacagatcc tgtctaggcc agagcggctg ctcttcatcc
     1981 tcgatggtgt agatgagcca ggatgggtct tgcaggagcc gagttctgag ctctgtctgc
     2041 actggagcca gccacagccg gcggatgcac tgctgggcag tttgctgggg aaaactatac
     2101 ttcccgaggc atccttcctg atcacggctc ggaccacagc tctgcagaac ctcattcctt
     2161 ctttggagca ggcacgttgg gtagaggtcc tggggttctc tgagtccagc aggaaggaat
     2221 atttctacag atatttcaca gatgaaaggc aagcaattag agcctttagg ttggtcaaat
     2281 caaacaaaga gctctgggcc ctgtgtcttg tgccctgggt gtcctggctg gcctgcactt
     2341 gcctgatgca gcagatgaag cggaaggaaa aactcacact gacttccaag accaccacaa
     2401 ccctctgtct acattacctt gcccaggctc tccaagctca gccattggga ccccagctca
     2461 gagacctctg ctctctggct gctgagggca tctggcaaaa aaagaccctt ttcagtccag
     2521 atgacctcag gaagcatggg ttagatgggg ccatcatctc caccttcttg aagatgggta
     2581 ttcttcaaga gcaccccatc cctctgagct acagcttcat tcacctctgt ttccaagagt
     2641 tctttgcagc aatgtcctat gtcttggagg atgagaaggg gagaggtaaa cattctaatt
     2701 gcatcataga tttggaaaag acgctagaag catatggaat acatggcctg tttggggcat
     2761 caaccacacg tttcctattg ggcctgttaa gtgatgaggg ggagagagag atggagaaca
     2821 tctttcactg ccggctgtct caggggagga acctgatgca gtgggtcccg tccctgcagc
     2881 tgctgctgca gccacactct ctggagtccc tccactgctt gtacgagact cggaacaaaa
     2941 cgttcctgac acaagtgatg gcccatttcg aagaaatggg catgtgtgta gaaacagaca
     3001 tggagctctt agtgtgcact ttctgcatta aattcagccg ccacgtgaag aagcttcagc
     3061 tgattgaggg caggcagcac agatcaacat ggagccccac catggtagtc ctgttcaggt
     3121 gggtcccagt cacagatgcc tattggcaga ttctcttctc cgtcctcaag gtcaccagaa
     3181 acctgaagga gctggaccta agtggaaact cgctgagcca ctctgcagtg aagagtcttt
     3241 gtaagaccct gagacgccct cgctgcctcc tggagaccct gcggttggct ggctgtggcc
     3301 tcacagctga ggactgcaag gaccttgcct ttgggctgag agccaaccag accctgaccg
     3361 agctggacct gagcttcaat gtgctcacgg atgctggagc caaacacctt tgccagagac
     3421 tgagacagcc gagctgcaag ctacagcgac tgcagctggt cagctgtggc ctcacgtctg
     3481 actgctgcca ggacctggcc tctgtgctta gtgccagccc cagcctgaag gagctagacc
     3541 tgcagcagaa caacctggat gacgttggcg tgcgactgct ctgtgagggg ctcaggcatc
     3601 ctgcctgcaa actcatacgc ctggggctgg accagacaac tctgagtgat gagatgaggc
     3661 aggaactgag ggccctggag caggagaaac ctcagctgct catcttcagc agacggaaac
     3721 caagtgtgat gacccctact gagggcctgg atacgggaga gatgagtaat agcacatcct
     3781 cactcaagcg gcagagactc ggatcagaga gggcggcttc ccatgttgct caggctaatc
     3841 tcaaactcct ggacgtgagc aagatcttcc caattgctga gattgcaggc aagagccacg
     3901 aggaaagctc cccagaggta gtaccggtgg aactcttgtg cgtgccttct cctgcctctc
     3961 aaggggacct gcatacgaag cctttgggga ctgacgatga cttctggggc cccacggggc
     4021 ctgtggctac tgaggtagtt gacaaagaaa agaacttgta ccgagttcac ttccctgtag
     4081 ctggctccta ccgctggccc aacacgggtc tctgctttgt gatgagagaa gcggtgaccg
     4141 ttgagattga attctgtgtg tgggaccagt tcctgggtga gatcaaccca cagcacagct
     4201 ggatggtggc agggcctctg ctggacatca aggctgagcc tggagctgtg gaagctgtgc
     4261 acctccctca ctttgtggct ctccaagggg gccatgtgga cacatccctg ttccaaatgg
     4321 cccactttaa agaggagggg atgctcctgg agaagccagc cagggtggag ctgcatcaca
     4381 tagttctgga aaaccccagc ttctccccct tgggagtcct cctgaaaatg atccataatg
     4441 ccctgcgctt cattcccgtc acctctgtgg tgttgcttta ccaccgcgtc catcctgagg
     4501 aagtcacctt ccacctctac ctgatcccaa gtgactgctc cattcggaag gccatagatg
     4561 atctagaaat gaaattccag tttgtgcgaa tccacaagcc acccccgctg accccacttt
     4621 atatgggctg tcgttacact gtgtctgggt ctggttcagg gatgctggaa atactcccca
     4681 aggaactgga gctctgctat cgaagccctg gagaagacca gctgttctcg gagttctacg
     4741 ttggccactt gggatcaggg atcaggctgc aagtgaaaga caagaaagat gagactctgg
     4801 tgtgggaggc cttggtgaaa ccaggaagga acaccagcca gccgtggaac ctcaggtgca
     4861 acagagacgc caggagatac ctcgagtcta gagactacaa agaccatgac ggtgattata
     4921 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     4981 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5041 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5101 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5161 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5221 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5281 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5341 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     5401 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5461 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5521 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     5581 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5641 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     5701 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5761 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5821 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     5881 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5941 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6001 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     6061 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6121 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6181 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6241 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6301 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     6361 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     6421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6481 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6541 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     6601 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6661 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     6721 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6781 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6841 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6901 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6961 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     7021 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     7081 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     7141 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     7201 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     7261 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     7321 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     7381 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7441 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7501 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     7561 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     7621 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     7681 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     7741 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7801 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     7861 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7921 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     8041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     8101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     8161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     8221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     8281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8461 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8701 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8881 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8941 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     9001 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     9061 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     9121 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     9181 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9241 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9301 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     9361 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9421 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9481 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9541 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9601 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9661 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9721 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9781 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9841 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9901 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9961 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    10021 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
    10081 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
    10141 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
    10201 cttagg
//
