LOCUS       V022798                 9737 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022798
VERSION     V022798
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             941..5227
                     /label="NLRP1(NM_014922)"
                     /note="NLRP1(NM_014922)"
                     /gene="NLRP1"
     CDS             5234..5299
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5337..5561
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5607..6035
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6049..6378
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6445..7239
                     /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    7413..7534
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7583..7599)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7607..7623
                     /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(7631..7661)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7676..7697
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7985..8570)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8741..9601)
                     /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(9602..9706)
                     /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 atccgccacc atggctggcg gagcctgggg
      961 ccgcctggcc tgttacttgg agttcctgaa gaaggaggag ctgaaggagt tccagcttct
     1021 gctcgccaat aaagcgcact ccaggagctc ttcgggtgag acacccgctc agccagagaa
     1081 gacgagtggc atggaggtgg cctcgtacct ggtggctcag tatggggagc agcgggcctg
     1141 ggacctagcc ctccatacct gggagcagat ggggctgagg tcactgtgcg cccaagccca
     1201 ggaaggggca ggccactctc cctcattccc ctacagccca agtgaacccc acctggggtc
     1261 tcccagccaa cccacctcca ccgcagtgct aatgccctgg atccatgaat tgccggcggg
     1321 gtgcacccag ggctcagaga gaagggtttt gagacagctg cctgacacat ctggacgccg
     1381 ctggagagaa atctctgcct cactcctcta ccaagctctt ccaagctccc cagaccatga
     1441 gtctccaagc caggagtcac ccaacgcccc cacatccaca gcagtgctgg ggagctgggg
     1501 atccccacct cagcccagcc tagcacccag agagcaggag gctcctggga cccaatggcc
     1561 tctggatgaa acgtcaggaa tttactacac agaaatcaga gaaagagaga gagagaaatc
     1621 agagaaaggc aggcccccat gggcagcggt ggtaggaacg cccccacagg cgcacaccag
     1681 cctacagccc caccaccacc catgggagcc ttctgtgaga gagagcctct gttccacatg
     1741 gccctggaaa aatgaggatt ttaaccaaaa attcacacag ctgctacttc tacaaagacc
     1801 tcaccccaga agccaagatc ccctggtcaa gagaagctgg cctgattatg tggaggagaa
     1861 tcgaggacat ttaattgaga tcagagactt atttggccca ggcctggata cccaagaacc
     1921 tcgcatagtc atactgcagg gggctgctgg aattgggaag tcaacactgg ccaggcaggt
     1981 gaaggaagcc tgggggagag gccagctgta tggggaccgc ttccagcatg tcttctactt
     2041 cagctgcaga gagctggccc agtccaaggt ggtgagtctc gctgagctca tcggaaaaga
     2101 tgggacagcc actccggctc ccattagaca gatcctgtct aggccagagc ggctgctctt
     2161 catcctcgat ggtgtagatg agccaggatg ggtcttgcag gagccgagtt ctgagctctg
     2221 tctgcactgg agccagccac agccggcgga tgcactgctg ggcagtttgc tggggaaaac
     2281 tatacttccc gaggcatcct tcctgatcac ggctcggacc acagctctgc agaacctcat
     2341 tccttctttg gagcaggcac gttgggtaga ggtcctgggg ttctctgagt ccagcaggaa
     2401 ggaatatttc tacagatatt tcacagatga aaggcaagca attagagcct ttaggttggt
     2461 caaatcaaac aaagagctct gggccctgtg tcttgtgccc tgggtgtcct ggctggcctg
     2521 cacttgcctg atgcagcaga tgaagcggaa ggaaaaactc acactgactt ccaagaccac
     2581 cacaaccctc tgtctacatt accttgccca ggctctccaa gctcagccat tgggacccca
     2641 gctcagagac ctctgctctc tggctgctga gggcatctgg caaaaaaaga cccttttcag
     2701 tccagatgac ctcaggaagc atgggttaga tggggccatc atctccacct tcttgaagat
     2761 gggtattctt caagagcacc ccatccctct gagctacagc ttcattcacc tctgtttcca
     2821 agagttcttt gcagcaatgt cctatgtctt ggaggatgag aaggggagag gtaaacattc
     2881 taattgcatc atagatttgg aaaagacgct agaagcatat ggaatacatg gcctgtttgg
     2941 ggcatcaacc acacgtttcc tattgggcct gttaagtgat gagggggaga gagagatgga
     3001 gaacatcttt cactgccggc tgtctcaggg gaggaacctg atgcagtggg tcccgtccct
     3061 gcagctgctg ctgcagccac actctctgga gtccctccac tgcttgtacg agactcggaa
     3121 caaaacgttc ctgacacaag tgatggccca tttcgaagaa atgggcatgt gtgtagaaac
     3181 agacatggag ctcttagtgt gcactttctg cattaaattc agccgccacg tgaagaagct
     3241 tcagctgatt gagggcaggc agcacagatc aacatggagc cccaccatgg tagtcctgtt
     3301 caggtgggtc ccagtcacag atgcctattg gcagattctc ttctccgtcc tcaaggtcac
     3361 cagaaacctg aaggagctgg acctaagtgg aaactcgctg agccactctg cagtgaagag
     3421 tctttgtaag accctgagac gccctcgctg cctcctggag accctgcggt tggctggctg
     3481 tggcctcaca gctgaggact gcaaggacct tgcctttggg ctgagagcca accagaccct
     3541 gaccgagctg gacctgagct tcaatgtgct cacggatgct ggagccaaac acctttgcca
     3601 gagactgaga cagccgagct gcaagctaca gcgactgcag ctggtcagct gtggcctcac
     3661 gtctgactgc tgccaggacc tggcctctgt gcttagtgcc agccccagcc tgaaggagct
     3721 agacctgcag cagaacaacc tggatgacgt tggcgtgcga ctgctctgtg aggggctcag
     3781 gcatcctgcc tgcaaactca tacgcctggg gctggaccag acaactctga gtgatgagat
     3841 gaggcaggaa ctgagggccc tggagcagga gaaacctcag ctgctcatct tcagcagacg
     3901 gaaaccaagt gtgatgaccc ctactgaggg cctggatacg ggagagatga gtaatagcac
     3961 atcctcactc aagcggcaga gactcggatc agagagggcg gcttcccatg ttgctcaggc
     4021 taatctcaaa ctcctggacg tgagcaagat cttcccaatt gctgagattg cagaggaaag
     4081 ctccccagag gtagtaccgg tggaactctt gtgcgtgcct tctcctgcct ctcaagggga
     4141 cctgcatacg aagcctttgg ggactgacga tgacttctgg ggccccacgg ggcctgtggc
     4201 tactgaggta gttgacaaag aaaagaactt gtaccgagtt cacttccctg tagctggctc
     4261 ctaccgctgg cccaacacgg gtctctgctt tgtgatgaga gaagcggtga ccgttgagat
     4321 tgaattctgt gtgtgggacc agttcctggg tgagatcaac ccacagcaca gctggatggt
     4381 ggcagggcct ctgctggaca tcaaggctga gcctggagct gtggaagctg tgcacctccc
     4441 tcactttgtg gctctccaag ggggccatgt ggacacatcc ctgttccaaa tggcccactt
     4501 taaagaggag gggatgctcc tggagaagcc agccagggtg gagctgcatc acatagttct
     4561 ggaaaacccc agcttctccc ccttgggagt cctcctgaaa atgatccata atgccctgcg
     4621 cttcattccc gtcacctctg tggtgttgct ttaccaccgc gtccatcctg aggaagtcac
     4681 cttccacctc tacctgatcc caagtgactg ctccattcgg aaggaactgg agctctgcta
     4741 tcgaagccct ggagaagacc agctgttctc ggagttctac gttggccact tgggatcagg
     4801 gatcaggctg caagtgaaag acaagaaaga tgagactctg gtgtgggagg ccttggtgaa
     4861 accaggagat ctcatgcctg caactactct gatccctcca gcccgcatag ccgtaccttc
     4921 acctctggat gccccgcagt tgctgcactt tgtggaccag tatcgagagc agctgatagc
     4981 ccgagtgaca tcggtggagg ttgtcttgga caaactgcat ggacaggtgc tgagccagga
     5041 gcagtacgag agggtgctgg ctgagaacac gaggcccagc cagatgcgga agctgttcag
     5101 cttgagccag tcctgggacc ggaagtgcaa agatggactc taccaagccc tgaaggagac
     5161 ccatcctcac ctcattatgg aactctggga gaagggcagc aaaaagggac tcctgccact
     5221 cagcagcctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     5281 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     5341 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5401 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5461 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5521 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     5581 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     5641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6001 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     6061 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6121 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6181 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6301 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6361 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     6421 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     6481 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     6541 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     6601 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6661 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6721 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6781 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6841 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6901 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6961 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     7021 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     7081 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     7141 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     7201 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     7261 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     7321 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     7381 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     7441 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7501 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     7561 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7621 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7681 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8401 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8521 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8581 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8641 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8701 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8761 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8821 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8881 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8941 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     9001 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     9061 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     9121 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     9181 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     9241 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     9301 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     9361 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9421 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9481 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9541 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9601 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9661 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9721 aagtgccacc tgacgtc
//
