LOCUS       sequence-423186-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gsdmd-NT 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
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..255
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     206..226
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             299..322
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1179..1801
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1378..1397)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1825..1845)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1843..2140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2194..2988
                     /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(2248..2267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2858..2877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3640..3774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3677..3696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3731..3750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3779..3798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3780..3798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3812..3828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3812..3828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3825..3847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3836..3852
                     /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(3860..3866,3867..3884,3885..3890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3905..3926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4043..4060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4214..4802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4294..4313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4973..5764,5765..5833))
                     /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     5596..5615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5834..5938)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5965..6420
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6052..6071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6262..6283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6546..6568
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6560..6577
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6561..6577
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        join(6738..7066,1..51)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
       61 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      121 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      181 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      241 tatataagca gagctcgttt agtgaaccgt cagaattaat tcagatcgat ctaccatgga
      301 ctacaaagac gatgacgaca agcttgcggc cgcaatgcca tcggcctttg agaaagtggt
      361 caagaatgtg atcaaggagg taagcggcag cagaggcgat ctcattccgg tggacagcct
      421 gcggaactcc accagcttca ggccctactg ccttctgaac aggaaatttt caagctcaag
      481 gttctggaaa ccccgttatt catgtgtcaa cctgtcaatc aaggacatcc tggagcccag
      541 tgctccagaa ccagaaccgg agtgttttgg ctccttcaaa gtctctgatg tcgtcgatgg
      601 gaacattcag ggcagagtga tgttgtcagg catgggagaa gggaaaattt ctggtggggc
      661 tgcagtgtct gacagttcca gtgcctccat gaatgtgtgt atactgcgtg tgactcagaa
      721 gacctgggag accatgcagc atgaaaggca ccttcagcag cctgagaaca aaatcctgca
      781 acagcttcgg agtcgtgggg atgacctgtt tgtggtgacc gaggtgctgc agacaaagga
      841 ggaagtgcag atcactgagg tccacagcca agagggctca ggccagttta cgctgcctgg
      901 agctttatgc ttgaagggtg aaggcaaggg ccaccaaagc cggaagaaga tggtgaccat
      961 tcctgcaggc agcatcctgg cattccgagt ggcccaactg cttattggct ctaaatggga
     1021 tatccttctc gtctcagatg agaaacagag gacctttgag ccctcctcag gtgacagaaa
     1081 agcagtgggc cagaggcacc atggcctcaa tgtgcttgct gcgctttgtt ccatcggaaa
     1141 gcagctcagt ctcctgtcag attagtctag aggatcccgg gtggcatccc tgtgacccct
     1201 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     1261 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     1321 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     1381 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     1441 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     1501 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     1561 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     1621 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     1681 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     1741 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     1801 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     1861 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     1921 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     1981 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2041 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2101 cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat cagcttggga
     2161 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2221 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2281 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2341 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     2401 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2461 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2521 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2581 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2641 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2701 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     2761 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     2821 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     2881 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     2941 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3001 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3061 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3121 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat cccctcgcga
     3181 gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg cagtgcaaat
     3241 ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc gaactgcagg
     3301 agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg cgcctgatac
     3361 agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc ctgaggtcac
     3421 tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca cccactcgcc
     3481 aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gccgatcata
     3541 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3601 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3661 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3721 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcc
     3781 ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt
     3841 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3961 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4021 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4081 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4141 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4621 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4921 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4981 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5041 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5101 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5161 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5221 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5281 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5341 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5401 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5461 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5521 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5581 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5641 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5701 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5761 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5821 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5881 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5941 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     6001 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     6061 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6121 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6181 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6241 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6301 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6361 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6421 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6481 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6541 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
     6601 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     6661 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     6721 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     6781 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     6841 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6901 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     6961 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
     7021 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgg
//
