LOCUS       sequence-260776-        7898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFP-tagged human NLRP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7898)
  AUTHORS   Khare S, Dorfleutner A, Bryan NB, Yun C, Radian AD, de Almeida L, 
            Rojanasakul Y, Stehlik C
  TITLE     An NLRP7-containing inflammasome mediates recognition of microbial 
            lipopeptides in human macrophages.
  JOURNAL   Immunity. 2012 Mar 23;36(3):464-76. doi: 
            10.1016/j.immuni.2012.02.001. Epub 2012 Feb 21.
  PUBMED    22361007
REFERENCE   2  (bases 1 to 7898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.immuni.2012.02.001"; journalName: "Immunity"; date: 
            "2012-03-23- 23"; volume: "36"; issue: "3"; pages: "464-76"
COMMENT     Sequence Label: pEGFP-C2-NLRP3
FEATURES             Location/Qualifiers
     source          1..7898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4626..4747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4663..4682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4717..4736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4754..5209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4891..4912)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5103..5122
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5236..5340
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5342..5699
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5342..5533
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5365..5385)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5550..5685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5734..6528
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5734..6528
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5788..5807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6398..6417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6716..6735)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6760..6807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7136..7724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7625..7644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atccgagatg aagatggcaa gcacccgctg caagctggcc
     1321 aggtacctgg aggacctgga ggatgtggac ttgaagaaat ttaagatgca cttagaggac
     1381 tatcctcccc agaagggctg catccccctc ccgaggggtc agacagagaa ggcagaccat
     1441 gtggatctag ccacgctaat gatcgacttc aatggggagg agaaggcgtg ggccatggcc
     1501 gtgtggatct tcgctgcgat caacaggaga gacctttatg agaaagcaaa aagagatgag
     1561 ccgaagtggg gttcagataa tgcacgtgtt tcgaatccca ctgtgatatg ccaggaagac
     1621 agcattgaag aggagtggat gggtttactg gagtaccttt cgagaatctc tatttgtaaa
     1681 atgaagaaag attaccgtaa gaagtacaga aagtacgtga gaagcagatt ccagtgcatt
     1741 gaagacagga atgcccgtct gggtgagagt gtgagcctca acaaacgcta cacacgactg
     1801 cgtctcatca aggagcaccg gagccagcag gagagggagc aggagcttct ggccatcggc
     1861 aagaccaaga cgtgtgagag ccccgtgagt cccattaaga tggagttgct gtttgacccc
     1921 gatgatgagc attctgagcc tgtgcacacc gtggtgttcc agggggcggc agggattggg
     1981 aaaacaatcc tggccaggaa gatgatgttg gactgggcat cggggacact ctaccaagac
     2041 aggtttgact atctgttcta tatccactgt cgggaggtga gccttgtgac acagaggagc
     2101 ctgggggacc tgatcatgag ctgctgcccc gacccaaacc cacccatcca caagatcgtg
     2161 agaaaaccct ccagaatcct cttcctcatg gacggcttcg atgagctgca aggtgccttt
     2221 gacgagcaca taggaccgct ctgcactgac tggcagaagg ccgagcgggg agacattctc
     2281 ctgagcagcc tcatcagaaa gaagctgctt cccgaggcct ctctgctcat caccacgaga
     2341 cctgtggccc tggagaaact gcagcacttg ctggaccatc ctcggcatgt ggagatcctg
     2401 ggtttctccg aggccaaaag gaaagagtac ttcttcaagt acttctctga tgaggcccaa
     2461 gccagggcag ccttcagtct gattcaggag aacgaggtcc tcttcaccat gtgcttcatc
     2521 cccctggtct gctggatcgt gtgcactgga ctgaaacagc agatggagag tggcaagagc
     2581 cttgcccaga catccaagac caccaccgcg gtgtacgtct tcttcctttc cagtttgctg
     2641 cagccccggg gagggagcca ggagcacggc ctctgcgccc acctctgggg gctctgctct
     2701 ttggctgcag atggaatctg gaaccagaaa atcctgtttg aggagtccga cctcaggaat
     2761 catggactgc agaaggcgga tgtgtctgct ttcctgagga tgaacctgtt ccaaaaggaa
     2821 gtggactgcg agaagttcta cagcttcatc cacatgactt tccaggagtt ctttgccgcc
     2881 atgtactacc tgctggaaga ggaaaaggaa ggaaggacga acgttccagg gagtcgtttg
     2941 aagcttccca gccgagacgt gacagtcctt ctggaaaact atggcaaatt cgaaaagggg
     3001 tatttgattt ttgttgtacg tttcctcttt ggcctggtaa accaggagag gacctcctac
     3061 ttggagaaga aattaagttg caagatctct cagcaaatca ggctggagct gctgaaatgg
     3121 attgaagtga aagccaaagc taaaaagctg cagatccagc ccagccagct ggaattgttc
     3181 tactgtttgt acgagatgca ggaggaggac ttcgtgcaaa gggccatgga ctatttcccc
     3241 aagattgaga tcaatctctc caccagaatg gaccacatgg tttcttcctt ttgcattgag
     3301 aactgtcatc gggtggagtc actgtccctg gggtttctcc ataacatgcc caaggaggaa
     3361 gaggaggagg aaaaggaagg ccgacacctt gatatggtgc agtgtgtcct cccaagctcc
     3421 tctcatgctg cctgttctca tggattggtg aacagccacc tcacttccag tttttgccgg
     3481 ggcctctttt cagttctgag caccagccag agtctaactg aattggacct cagtgacaat
     3541 tctctggggg acccagggat gagagtgttg tgtgaaacgc tccagcatcc tggctgtaac
     3601 attcggagat tgtggttggg gcgctgtggc ctctcgcatg agtgctgctt cgacatctcc
     3661 ttggtcctca gcagcaacca gaagctggtg gagctggacc tgagtgacaa cgccctcggt
     3721 gacttcggaa tcagacttct gtgtgtggga ctgaagcacc tgttgtgcaa tctgaagaag
     3781 ctctggttgg tcagctgctg cctcacatca gcatgttgtc aggatcttgc atcagtattg
     3841 agcaccagcc attccctgac cagactctat gtgggggaga atgccttggg agactcagga
     3901 gtcgcaattt tatgtgaaaa agccaagaat ccacagtgta acctgcagaa actggggttg
     3961 gtgaattctg gccttacgtc agtctgttgt tcagctttgt cctcggtact cagcactaat
     4021 cagaatctca cgcaccttta cctgcgaggc aacactctcg gagacaaggg gatcaaacta
     4081 ctctgtgagg gactcttgca ccccgactgc aagcttcagg tgttggaatt agacaactgc
     4141 aacctcacgt cacactgctg ctgggatctt tccacacttc tgacctccag ccagagcctg
     4201 cgaaagctga gcctgggcaa caatgacctg ggcgacctgg gggtcatgat gttctgtgaa
     4261 gtgctgaaac agcagagctg cctcctgcag aacctggggt tgtctgaaat gtatttcaat
     4321 tatgagacaa aaagtgcgtt agaaacactt caagaagaaa agcctgagct gaccgtcgtc
     4381 tttgagcctt cttggtagga gtggaaacgg ggctgccaga cgccagtgtt ctccggtccc
     4441 tccagctggg ggccctcagg tggagagagc tgcgactcga cggtaccgcg ggcccgggat
     4501 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     4561 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4621 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4681 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4741 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4801 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4861 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4921 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4981 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5041 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5101 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5161 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5221 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5281 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5341 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5401 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5521 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5581 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5641 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     5701 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5761 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5821 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5881 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     5941 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6001 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6061 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6121 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6181 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6241 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     6301 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6361 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6421 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6481 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6541 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6601 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6661 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     6721 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6781 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     6841 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     6901 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6961 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     7021 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7081 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7741 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7801 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     7861 acggggtcat tagttcatag cccatatatg gagttccg
//
