LOCUS       V040615                 9206 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040615
VERSION     V040615
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..4726
                     /label="Nlrp3(NM_001359638)"
                     /note="Nlrp3(NM_001359638)"
                     /gene="Nlrp3"
     polyA_signal    4773..4997
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5043..5471
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5485..5814
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5881..6675
                     /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    6849..6970
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7019..7035)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7043..7059
                     /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(7067..7097)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7112..7133
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7421..8006)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8177..9037)
                     /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(9038..9142)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg acgagtgtcc gttgcaagct ggctcagtat ctagaggacc ttgaagatgt
     1681 ggacctcaag aaattcaaaa tgcatttgga agattacccg cccgagaaag gctgtatccc
     1741 agtccccagg ggccagatgg agaaggcaga tcacttggat ctagccacac tcatgattga
     1801 cttcaatggc gaggagaagg cctgggccat ggctgtgtgg atctttgctg cgatcaacag
     1861 gcgagacctc tgggaaaaag ctaagaagga ccagccagag tggaatgaca cgtgtacatc
     1921 acattcctct atggtatgcc aggaggacag ccttgaagaa gagtggatgg gtttgctggg
     1981 atatctctcc cgcatctcca tttgtaaaaa gaagaaagat tactgtaaga tgtacagacg
     2041 acatgtgaga agcaggttct actctatcaa ggacaggaac gcgcgtctag gtgagagtgt
     2101 ggacctcaac agtcgctaca cgcagctcca actggtcaag gagcatccaa gcaagcagga
     2161 gcgggagcat gaactcctga ccatcggccg gactaaaatg cgggacagcc ccatgagttc
     2221 ccttaagctg gagctgctgt ttgagcccga ggacgggcac tcggagcctg tgcacacagt
     2281 ggtgttccag ggagcagcag gcatcgggaa aaccatccta gccaggaaga ttatgttgga
     2341 ctgggcactg ggaaagctct tcaaagacaa atttgactat ttgttcttta tccactgccg
     2401 agaggtgagc ctcaggacgc caaggagtct agcagacctg attgtcagct gctggcctga
     2461 cccaaaccca ccagtgtgca agatcctgcg caagccttcc aggatcctct tcctcatgga
     2521 tggctttgat gagctacaag gggcctttga cgagcacatt ggggaggtct gcacagactg
     2581 gcaaaaggct gtgcggggag acattctgct aagcagcctc atccgaaaga aactgctgcc
     2641 caaggcctct ctgctcataa cgacgaggcc ggtagccttg gagaaactgc agcatctcct
     2701 ggaccacccc cgccatgtgg agatcctagg tttctctgag gccaaaagga aggagtattt
     2761 ctttaagtat ttctccaacg agctgcaggc ccgggaggcc ttcaggctga tccaagagaa
     2821 tgaggtcctc tttaccatgt gcttcatccc cctggtctgc tggattgtgt gcacggggct
     2881 aaagcaacag atggagaccg ggaagagcct ggcccagacc tccaagacca ctacggccgt
     2941 ctacgtcttc ttcctttcca gcctgctgca atcccggggg ggcattgagg agcatctctt
     3001 ctctgactac ctacaggggc tctgttcact ggctgcggat ggaatttgga accagaaaat
     3061 cctatttgag gagtgtgatc tgcggaagca cggcctgcag aagactgacg tctccgcttt
     3121 cctgaggatg aacgtgttcc agaaggaagt ggactgcgag agattctaca gcttcagcca
     3181 catgactttc caggagttct tcgctgctat gtactatttg ctggaagagg aggcagaggg
     3241 ggagaccgtg aggaaaggac caggaggttg ttcagatctt ctgaaccgag acgtgaaggt
     3301 cctactagaa aattacggca agtttgaaaa aggctatctg atttttgttg tccgattcct
     3361 ctttggcctt gtaaaccagg agagaacctc ttatttggag aagaaactaa gttgcaagat
     3421 ctctcagcaa gtcagactgg aactactgaa gtggattgaa gtgaaagcca aggccaagaa
     3481 gctgcagtgg cagcccagcc aactggaact gttctactgc ctgtacgaga tgcaggagga
     3541 agactttgtg cagagtgcca tggaccactt tcccaaaatt gagatcaacc tctctaccag
     3601 aatggaccac gtggtttcct ccttttgtat taagaactgt catagggtca aaacgctttc
     3661 cctgggtttt tttcacaact cgcccaagga ggaagaagaa gagaggagag gaggtcgacc
     3721 cttggaccag gttcagtgtg ttttcccaga cactcatgtt gcctgttctt ccagactggt
     3781 gaactgctgc ctcacttcta gcttctgccg tggtctcttc tcaagtctaa gcaccaaccg
     3841 gagcctcact gaactggacc tcagtgacaa tactctggga gacccgggca tgagggtgct
     3901 gtgtgaggca ctccagcacc caggctgtaa cattcagaga ctgtggttgg ggcgctgcgg
     3961 actgtcccat caatgctgct tcgacatctc ctctgtcctg agcagcagcc agaagctggt
     4021 ggagctggac ctcagtgaca atgccctggg ggactttgga atcagattgc tgtgtgtggg
     4081 actgaagcac ctgctctgca acctccagaa actgtggttg gtgagctgct gtctcacatc
     4141 cgcgtgttgt caggatctcg cattggttct gagctccaac cattctctga ccagactgta
     4201 cattggagaa aatgccttgg gagactcagg agtccaagtt ttgtgtgaaa agatgaagga
     4261 cccacagtgt aacttgcaga agctggggtt ggtgaattcc ggccttactt caatctgttg
     4321 ttcagctctg acctctgtgc tcaaaaccaa ccagaacttc acacacctct atctacgaag
     4381 caatgccctt ggagacacag gactcaggct cctctgtgag gggcttctgc acccggactg
     4441 taaactacag atgctggaat tagacaactg cagcctcacc tcacacagct gctggaatct
     4501 ctccacaatt ctgacccaca accacagcct tcggaagctg aacctgggca acaatgatct
     4561 tggcgatctg tgcgtggtga ccctctgtga ggtgctgaaa cagcagggct gcctcctgca
     4621 gagcctacag ttgggtgaaa tgtacttaaa tcgtgaaaca aaacgtgcct tagaagcgct
     4681 ccaggaagaa aagcctgagc tgactatagt cttcgagatt tcctggtagg gatcctctag
     4741 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4801 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4861 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4921 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4981 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     5041 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5101 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5161 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5221 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5281 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5341 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5401 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5461 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5641 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5701 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5761 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5821 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5881 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5941 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6001 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6061 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6121 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6181 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6241 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6301 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6361 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6421 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6481 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6541 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6601 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6661 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6721 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6781 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6841 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6901 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6961 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     7021 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7081 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7141 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7201 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7261 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7321 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7921 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     7981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8461 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8581 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8701 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8761 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8821 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8881 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8941 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9001 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9061 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9121 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     9181 cgggagatct cccgatcccc tatggt
//
