LOCUS       V027203                 9057 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027203
VERSION     V027203
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..3737
                     /label="NLRP4(NM_134444)"
                     /note="NLRP4(NM_134444)"
                     /gene="NLRP4"
     CDS             3744..3809
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3833..4421
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4487..4535
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4737..5165
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5179..5508
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5575..6369
                     /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    6545..6666
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6715..6731)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6739..6755
                     /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(6763..6793)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6808..6829
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7117..7705)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7876..8736)
                     /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(8737..8841)
                     /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 ccaccatggc agcctctttc ttctctgatt
      781 ttggtcttat gtggtatctg gaggagctca aaaaggagga gttcaggaaa tttaaagaac
      841 atctcaagca aatgactttg cagcttgaac tcaagcagat tccctggact gaggtcaaaa
      901 aagcatcccg ggaagaactt gcaaacctct tgatcaagca ctatgaagaa caacaagctt
      961 ggaacataac cttaagaatc tttcaaaaga tggatagaaa ggatctctgc atgaaggtca
     1021 tgagggagag aacaggatac acaaagacct atcaagctca cgcaaagcag aaattcagcc
     1081 gcttatggtc cagcaagtct gtcactgaga ttcacctata ctttgaggag gaagtcaagc
     1141 aagaagaatg tgaccatttg gaccgccttt ttgctcccaa ggaagctggg aaacagccac
     1201 gtacagtgat cattcaagga ccacaaggaa ttggaaaaac gacactcctg atgaagctga
     1261 tgatggcctg gtcggacaac aagatctttc gggataggtt cctgtacacg ttctatttct
     1321 gctgcagaga actgagggag ttgccgccaa cgagtttggc tgacttgatt tccagagagt
     1381 ggcctgaccc cgctgctcct ataacagaga tcgtgtctca accggagaga ctcttgttcg
     1441 tcatcgacag cttcgaagag ctgcagggcg gcttgaacga acccgattcg gatctgtgtg
     1501 gtgacttgat ggagaaacgg ccggtgcagg tgcttctgag cagtttgctg aggaagaaga
     1561 tgctcccgga ggcctccctg ctcatcgcta tcaaacccgt gtgcccgaag gagctccggg
     1621 atcaggtgac gatctcagaa atctaccagc cccggggatt caacgagagt gataggttag
     1681 tgtatttctg ctgtttcttc aaagacccga aaagagccat ggaagccttc aatcttgtaa
     1741 gagaaagtga acagctgttt tccatatgcc aaatcccgct cctctgctgg atcctgtgta
     1801 ccagtctgaa gcaagagatg cagaaaggaa aagacctggc cctgacctgc cagagcacta
     1861 cctctgtgta ctcctctttc gtctttaacc tgttcacacc tgagggtgcc gagggcccga
     1921 ctccgcaaac ccagcaccag ctgaaggccc tgtgctccct ggctgcagag ggtatgtgga
     1981 cagacacatt tgagttttgt gaagacgacc tccggagaaa tggggttgtt gacgctgaca
     2041 tccctgcgct gctgggcacc aagatacttc tgaagtacgg ggagcgtgag agctcctacg
     2101 tgttcctcca cgtgtgtatc caggagttct gtgccgcctt gttctatttg ctcaagagcc
     2161 accttgatca tcctcaccca gctgtgagat gtgtacagga attgctagtt gccaattttg
     2221 aaaaagcaag gagagcacat tggatttttt tggggtgttt tctaactggc cttttaaata
     2281 aaaaggaaca agaaaaactg gatgcgtttt ttggcttcca actgtcccaa gagataaagc
     2341 agcaaattca ccagtgcctg aagagcttag gggagcgtgg caatcctcag ggacaggtgg
     2401 attccttggc gatattttac tgtctctttg aaatgcagga tcctgccttt gtgaagcagg
     2461 cagtgaacct cctccaagaa gctaactttc atattattga caacgtggac ttggtggttt
     2521 ctgcctactg cttaaaatac tgctccagct tgaggaaact ctgtttttcc gttcaaaatg
     2581 tctttaagaa agaggatgaa cacagctcta cgtcggatta cagcctcatc tgttggcatc
     2641 acatctgctc tgtgctcacc accagcgggc acctcagaga gctccaggtg caggacagca
     2701 ccctcagcga gtcgaccttt gtgacctggt gtaaccagct gaggcatccc agctgtcgcc
     2761 ttcagaagct tggaataaat aacgtttcct tttctggcca gagtgttctg ctctttgagg
     2821 tgctctttta tcagccagac ttgaaatacc tgagcttcac cctcacgaaa ctctctcgtg
     2881 atgacatcag gtccctctgt gatgccttga actacccagc aggcaacgtc aaagagctag
     2941 cgctggtaaa ttgtcacctc tcacccattg attgtgaagt ccttgctggc cttctaacca
     3001 acaacaagaa gctgacgtat ctgaatgtat cctgcaacca gttagacaca ggcgtgcccc
     3061 ttttgtgtga agccctgtgc agcccagaca cggtcctggt atacctgatg ttggctttct
     3121 gccacctcag cgagcagtgc tgcgaataca tctctgaaat gcttctgcgt aacaagagcg
     3181 tgcgctatct agacctcagt gccaatgtcc tgaaggacga aggactgaaa actctctgcg
     3241 aggccttgaa acatccggac tgctgcctgg attcactgtg tttggtaaaa tgttttatca
     3301 ctgctgctgg ctgtgaagac ctcgcctctg ctctcatcag caatcaaaac ctgaagattc
     3361 tgcaaattgg gtgcaatgaa atcggagatg tgggtgtgca gctgttgtgt cgggctctga
     3421 cgcatacgga ttgccgctta gagattcttg ggttggaaga atgtgggtta acgagcacct
     3481 gctgtaagga tctcgcgtct gttctcacct gcagtaagac cctgcagcag ctcaacctga
     3541 ccttgaacac cttggaccac acaggggtgg ttgtactctg tgaggccctg agacacccag
     3601 agtgtgccct gcaggtgctc gggctgagaa aaactgattt tgatgaggaa acccaggcac
     3661 ttctgacggc tgaggaagag agaaatccta acctgaccat cacagacgac tgtgacacaa
     3721 tcacaagggt agagatcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     3781 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     3841 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3901 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3961 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4021 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4081 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4141 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4201 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4261 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4321 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4381 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     4441 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4501 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4561 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4621 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4681 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     4741 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4801 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4861 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     4921 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4981 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5041 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5101 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5161 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5341 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5401 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5461 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5521 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5581 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5641 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5701 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5761 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5821 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5881 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5941 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6001 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6061 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6121 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6181 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6241 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6301 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6361 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     6421 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6481 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6541 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6601 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6661 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7801 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7861 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7921 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7981 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8041 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8101 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8161 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8221 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8281 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8341 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8401 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8461 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8521 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8581 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8641 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8701 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8761 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8821 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8881 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     8941 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     9001 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
