LOCUS       sequence-436177-        6761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gasdermin-D_29-484 is a non-functional GSDMD formed by SARS-CoV-2 
            3CLpro cleavage in the N-terminal domain. Transfection in HEK293 
            cells showed no pore-forming activity or induction of pyroptosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6761)
  AUTHORS   Grin PM, Baid K, de Jesus HCR, Kozarac N, Bell PA, Jiang SZ, 
            Kappelhoff R, Butler GS, Leborgne NGF, Pan C, Pablos I, Machado Y, 
            Vederas JC, Kim H, Benarafa C, Banerjee A, Overall CM
  TITLE     SARS-CoV-2 3CL(pro) (main protease) regulates caspase activation of 
            gasdermin-D/E pores leading to secretion and extracellular activity 
            of 3CL(pro).
  JOURNAL   Cell Rep. 2024 Dec 24;43(12):115080. doi: 
            10.1016/j.celrep.2024.115080. Epub 2024 Dec 12.
  PUBMED    39673710
REFERENCE   2  (bases 1 to 6761)
  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"; journalName: "Cell Rep.";
            date: "2024-12-24"; volume: "43(12):115080. doi"; pages: " 
            10.1016/j.celrep.2024.115080. Epub 2024 Dec 12"
COMMENT     Sequence Label: pcDNA3.1-GSDMD_29-484
FEATURES             Location/Qualifiers
     source          1..6761
                     /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             2309..2332
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2352..2369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2358..2582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2628..3056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2715..2734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2925..2946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3065..3085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3070..3399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3250..3385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3312..3331
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3466..4260
                     /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(3520..3539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4130..4149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4434..4555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4471..4490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4525..4544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4604..4620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4604..4620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4617..4639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4628..4644
                     /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(4652..4658,4659..4676,4677..4682))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4697..4718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4835..4852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5006..5594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5086..5105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5765..6556,6557..6625))
                     /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     6388..6407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6626..6730)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgagctcca ctggcttcca
      961 gccctactgc ctggtggtta ggaagccctc aagctcatgg ttctggaaac cccgttataa
     1021 gtgtgtcaac ctgtctatca aggacatcct ggagccggat gccgcggaac cagacgtgca
     1081 gcgtggcagg agcttccact tctacgatgc catggatggg cagatacagg gcagcgtgga
     1141 gctggcagcc ccaggacagg caaagatcgc aggcggggcc gcggtgtctg acagctccag
     1201 cacctcaatg aatgtgtact cgctgagtgt ggaccctaac acctggcaga ctctgctcca
     1261 tgagaggcac ctgcggcagc cagaacacaa agtcctgcag cagctgcgca gccgcgggga
     1321 caacgtgtac gtggtgactg aggtgctgca gacacagaag gaggtggaag tcacgcgcac
     1381 ccacaagcgg gagggctcgg gccggttttc cctgcccgga gccacgtgct tgcagggtga
     1441 gggccagggc catctgagcc agaagaagac ggtcaccatc ccctcaggca gcaccctcgc
     1501 attccgggtg gcccagctgg ttattgactc tgacttggac gtccttctct tcccggataa
     1561 gaagcagagg accttccagc cacccgcgac aggccacaag cgttccacga gcgaaggcgc
     1621 ctggccacag ctgccctctg gcctctccat gatgaggtgc ctccacaact tcctgacaga
     1681 tggggtccct gcggaggggg cgttcactga agacttccag ggcctacggg cagaggtgga
     1741 gaccatctcc aaggaactgg agcttttgga cagagagctg tgccagctgc tgctggaggg
     1801 cctggagggg gtgctgcggg accagctggc cctgcgagcc ttggaggagg cgctggagca
     1861 gggccagagc cttgggccgg tggagcccct ggacggtcca gcaggtgctg tcctggagtg
     1921 cctggtgttg tcctccggaa tgctggtgcc ggaactcgct atccctgttg tctacctgct
     1981 gggggcactg accatgctga gtgaaacgca gcacaagctg ctggcggagg cgctggagtc
     2041 gcagaccctg ttggggccgc tcgagctggt gggcagcctc ttggagcaga gtgccccgtg
     2101 gcaggagcgc agcaccatgt ccctgccccc cgggctcctg gggaacagct ggggcgaagg
     2161 agcaccggcc tgggtcttgc tggacgagtg tggcctagag ctgggggagg acactcccca
     2221 cgtgtgctgg gagccgcagg cccagggccg catgtgtgca ctctacgcct ccctggcact
     2281 gctatcagga ctgagccagg agccccacga ttacaaggat gacgacgata agtgataaac
     2341 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2401 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2461 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2521 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2581 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2641 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2701 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2761 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2821 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2941 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3001 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3301 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3421 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3481 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3541 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3601 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3661 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3721 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3781 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3841 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3901 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3961 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4021 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4081 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4141 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4201 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4261 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4321 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4381 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4441 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4501 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4561 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4621 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4681 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4741 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4801 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4861 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4921 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4981 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5041 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5341 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5401 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5461 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5521 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5581 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5641 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5701 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5761 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5821 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5881 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5941 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6001 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6061 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6121 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6181 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6241 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6301 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6361 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6421 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6481 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6541 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6601 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6661 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6721 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
