LOCUS       sequence-266234-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gsdmd-NT mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  AUTHORS   Liu X, Zhang Z, Ruan J, Pan Y, Magupalli VG, Wu H, Lieberman J
  TITLE     Inflammasome-activated gasdermin D causes pyroptosis by forming 
            membrane pores.
  JOURNAL   Nature. 2016 Jul 6;535(7610):153-8. doi: 10.1038/nature18629.
  PUBMED    27383986
REFERENCE   2  (bases 1 to 7066)
  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"; journalName: 
            "Nature"; date: "2016-07-6- 6"; volume: "535"; issue: "7610"; pages:
            "153-8"
COMMENT     Sequence Label: Flag-Gsdmd-NT C265A
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..42
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     34..51
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     35..51
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        212..591
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        592..795
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     746..766
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             839..862
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1719..2341
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1918..1937)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2365..2385)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2383..2680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2734..3528
                     /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(2788..2807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3398..3417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4180..4314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4217..4236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4271..4290
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4319..4338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4320..4338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4352..4368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4352..4368)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4365..4387)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4376..4392
                     /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(4400..4406,4407..4424,4425..4430))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4445..4466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4583..4600)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4754..5342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4834..4853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5513..6304,6305..6373))
                     /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     6136..6155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6374..6478)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6505..6960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6592..6611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6802..6823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
       61 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
      121 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      181 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
      241 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      301 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      361 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      421 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      481 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      541 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      601 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      661 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      721 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      781 tatataagca gagctcgttt agtgaaccgt cagaattaat tcagatcgat ctaccatgga
      841 ctacaaagac gatgacgaca agcttgcggc cgcaatgcca tcggcctttg agaaagtggt
      901 caagaatgtg atcaaggagg taagcggcag cagaggcgat ctcattccgg tggacagcct
      961 gcggaactcc accagcttca ggccctactg ccttctgaac aggaaatttt caagctcaag
     1021 gttctggaaa ccccgttatt catgtgtcaa cctgtcaatc aaggacatcc tggagcccag
     1081 tgctccagaa ccagaaccgg agtgttttgg ctccttcaaa gtctctgatg tcgtcgatgg
     1141 gaacattcag ggcagagtga tgttgtcagg catgggagaa gggaaaattt ctggtggggc
     1201 tgcagtgtct gacagttcca gtgcctccat gaatgtgtgt atactgcgtg tgactcagaa
     1261 gacctgggag accatgcagc atgaaaggca ccttcagcag cctgagaaca aaatcctgca
     1321 acagcttcgg agtcgtgggg atgacctgtt tgtggtgacc gaggtgctgc agacaaagga
     1381 ggaagtgcag atcactgagg tccacagcca agagggctca ggccagttta cgctgcctgg
     1441 agctttatgc ttgaagggtg aaggcaaggg ccaccaaagc cggaagaaga tggtgaccat
     1501 tcctgcaggc agcatcctgg cattccgagt ggcccaactg cttattggct ctaaatggga
     1561 tatccttctc gtctcagatg agaaacagag gacctttgag ccctcctcag gtgacagaaa
     1621 agcagtgggc cagaggcacc atggcctcaa tgtgcttgct gcgcttgctt ccatcggaaa
     1681 gcagctcagt ctcctgtcag attagtctag aggatcccgg gtggcatccc tgtgacccct
     1741 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     1801 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     1861 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     1921 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     1981 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2041 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2101 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2161 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     2221 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     2281 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     2341 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2401 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2521 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2581 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2641 cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat cagcttggga
     2701 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2761 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2821 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2881 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     2941 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3001 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3061 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3121 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3181 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3241 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3301 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3361 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3421 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3481 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3541 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3601 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3661 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat cccctcgcga
     3721 gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg cagtgcaaat
     3781 ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc gaactgcagg
     3841 agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg cgcctgatac
     3901 agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc ctgaggtcac
     3961 tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca cccactcgcc
     4021 aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gccgatcata
     4081 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4141 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4201 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4261 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcc
     4321 ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt
     4381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6481 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6541 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     6601 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6661 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6721 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6781 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6841 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6901 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6961 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     7021 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagt
//
