LOCUS       sequence-142337-        7311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal Flag-tagged amino acids 91-710 of NLRP3 (the 
            nucleotide-binding domain) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7311)
  AUTHORS   Shi H, Wang Y, Li X, Zhan X, Tang M, Fina M, Su L, Pratt D, Bu CH, 
            Hildebrand S, Lyon S, Scott L, Quan J, Sun Q, Russell J, Arnett S, 
            Jurek P, Chen D, Kravchenko VV, Mathison JC, Moresco EM, Monson NL, 
            Ulevitch RJ, Beutler B
  TITLE     NLRP3 activation and mitosis are mutually exclusive events 
            coordinated by NEK7, a new inflammasome component.
  JOURNAL   Nat Immunol. 2016 Mar;17(3):250-8. doi: 10.1038/ni.3333. Epub 2015 
            Dec 7.
  PUBMED    26642356
REFERENCE   2  (bases 1 to 7311)
  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: "10.1038/ni.3333"; 
            journalName: "Nat Immunol"; date: "2016-03"; volume: "17"; issue: 
            "3"; pages: "250-8"
COMMENT     Sequence Label: pcDNA3-N-Flag-NLRP3 91-710
FEATURES             Location/Qualifiers
     source          1..7311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             922..945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2863..2881)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2864..2881)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2901..2918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2907..3131
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3177..3605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3264..3283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3474..3495
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3614..3634)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3619..3949
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3800..3935
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3862..3881
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4016..4810
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4070..4089)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4680..4699
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4984..5105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5021..5040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5075..5094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5154..5170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5154..5170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5167..5189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5178..5194
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5202..5208,5209..5226,5227..5232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5247..5268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5385..5402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5556..6144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5636..5655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6315..7106,7107..7175))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6938..6957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7176..7280)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat cgcccaccat ggactacaaa gacgatgacg ataaaatcga tatcgaattc
      961 atgagtggaa tgacacgtgt acatcacatt cctctatggt atgccaggag gacagccttg
     1021 aagaagagtg gatgggtttg ctgggatatc tctcccgcat ctccatttgt aaaaagaaga
     1081 aagattactg taagatgtac agacgacatg tgagaagcag gttctactct atcaaggaca
     1141 ggaacgcgcg tctaggtgag agtgtggacc tcaacagtcg ctacacgcag ctccaactgg
     1201 tcaaggagca tccaagcaag caggagcggg agcatgaact cctgaccatc ggccggacta
     1261 aaatgcggga cagccccatg agttccctta agctggagct gctgtttgag cccgaggacg
     1321 ggcactcgga gcctgtgcac acagtggtgt tccagggagc agcaggcatc gggaaaacca
     1381 tcctagccag gaagattatg ttggactggg cactgggaaa gctcttcaaa gacaaatttg
     1441 actatttgtt ctttatccac tgccgagagg tgagcctcag gacgccaagg agtctagcag
     1501 acctgattgt cagctgctgg cctgacccaa acccaccagt gtgcaagatc ctgcgcaagc
     1561 cttccaggat cctcttcctc atggatggct ttgatgagct acaaggggcc tttgacgagc
     1621 acattgggga ggtctgcaca gactggcaaa aggctgtgcg gggagacatt ctgctaagca
     1681 gcctcatccg aaagaaactg ctgcccaagg cctctctgct cataacgacg aggccggtag
     1741 ccttggagaa actgcagcat ctcctggacc acccccgcca tgtggagatc ctaggtttct
     1801 ctgaggccaa aaggaaggag tatttcttta agtatttctc caacgagctg caggcccggg
     1861 aggccttcag gctgatccaa gagaatgagg tcctctttac catgtgcttc atccccctgg
     1921 tctgctggat tgtgtgcacg gggctaaagc aacagatgga gaccgggaag agcctggccc
     1981 agacctccaa gaccactacg gccgtctacg tcttcttcct ttccagcctg ctgcaatccc
     2041 gggggggcat tgaggagcat ctcttctctg actacctaca ggggctctgt tcactggctg
     2101 cggatggaat ttggaaccag aaaatcctat ttgaggagtg tgatctgcgg aagcacggcc
     2161 tgcagaagac tgacgtctcc gctttcctga ggatgaacgt gttccagaag gaagtggact
     2221 gcgagagatt ctacagcttc agccacatga ctttccagga gttcttcgct gctatgtact
     2281 atttgctgga agaggaggca gagggggaga ccgtgaggaa aggaccagga ggttgttcag
     2341 atcttctgaa ccgagacgtg aaggtcctac tagaaaatta cggcaagttt gaaaaaggct
     2401 atctgatttt tgttgtccga ttcctctttg gccttgtaaa ccaggagaga acctcttatt
     2461 tggagaagaa actaagttgc aagatctctc agcaagtcag actggaacta ctgaagtgga
     2521 ttgaagtgaa agccaaggcc aagaagctgc agtggcagcc cagccaactg gaactgttct
     2581 actgcctgta cgagatgcag gaggaagact ttgtgcagag tgccatggac cactttccca
     2641 aaattgagat caacctctct accagaatgg accacgtggt ttcctccttt tgtattaaga
     2701 actgtcatag ggtcaaaacg ctttccctgg gtttttttca caactcgccc aaggaggaag
     2761 aagaagagag gagaggaggt cgacccttgg accaggttca gtgtgttttc ccagacactc
     2821 atgttgcctg tttagctcga gcatgcatct agagggccct attctatagt gtcacctaaa
     2881 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2941 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3001 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3061 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3121 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3181 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3241 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3301 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3361 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3421 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3481 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3541 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3601 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc
     3661 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3721 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3781 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3841 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3901 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3961 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4021 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4081 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4141 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4201 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4261 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4321 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4381 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4441 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4501 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4561 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4621 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4681 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4741 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4801 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4861 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4921 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4981 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5041 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5101 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5521 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     5761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6241 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6301 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6361 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6421 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6481 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6541 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6601 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6661 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6721 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6781 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6841 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6901 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6961 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7021 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7081 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7141 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7201 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7261 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
