LOCUS       sequence-394032-        7935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GSDMD gene in mammalian cells by retroviral 
            transduction .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7935)
  AUTHORS   Devant P, Borsic E, Ngwa EM, Xiao H, Chouchani ET, Thiagarajah JR, 
            Hafner-Bratkovic I, Evavold CL, Kagan JC
  TITLE     Gasdermin D pore-forming activity is redox-sensitive.
  JOURNAL   Cell Rep. 2023 Jan 31;42(1):112008. doi: 
            10.1016/j.celrep.2023.112008. Epub 2023 Jan 19.
  PUBMED    36662620
REFERENCE   2  (bases 1 to 7935)
  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: "2023-01-31"; volume: "42(1):112008. doi"; pages: " 
            10.1016/j.celrep.2023.112008. Epub 2023 Jan 19"
COMMENT     Sequence Label: pMSCV-GSDMD-Myc
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2904..2933
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2968..3534
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2984..3536
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3128..3145)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3355..3374
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3548..3550,3551..3553,3554..4267)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3593..3614)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3854..3873)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4201..4222
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4329..4843
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5005..5021
                     /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(5029..5035,5036..5053,5054..5059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5074..5095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5212..5229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5383..5971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5463..5482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6142..6933,6934..7002))
                     /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     6765..6784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7003..7107)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7175..7193
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7231..7253)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7353..7372
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7566..7588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7716..7735
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7935,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg agatcgatgc ggccgcgcca
     1441 ccatgccatc ggcctttgag aaagtggtca agaatgtgat caaggaggta agcggcagca
     1501 gaggcgatct cattccggtg gacagcctgc ggaactccac cagcttcagg ccctactgcc
     1561 ttctgaacag gaaattttca agctcaaggt tctggaaacc ccgttattca tgtgtcaacc
     1621 tgtcaatcaa ggacatcctg gagcccagtg ctccagaacc agaaccggag tgttttggct
     1681 ccttcaaagt ctctgatgtc gtcgatggga acattcaggg cagagtgatg ttgtcaggca
     1741 tgggagaagg gaaaatttct ggtggggctg cagtgtctga cagttccagt gcctccatga
     1801 atgtgtgtat actgcgtgtg actcagaaga cctgggagac catgcagcat gaaaggcacc
     1861 ttcagcagcc tgagaacaaa atcctgcaac agcttcggag tcgtggggat gacctgtttg
     1921 tggtgaccga ggtgctgcag acaaaggagg aagtgcagat cactgaggtc cacagccaag
     1981 agggctcagg ccagtttacg ctgcctggag ctttatgctt gaagggtgaa ggcaagggcc
     2041 accaaagccg gaagaagatg gtgaccattc ctgcaggcag catcctggca ttccgagtgg
     2101 cccaactgct tattggctct aaatgggata tccttctcgt ctcagatgag aaacagagga
     2161 cctttgagcc ctcctcaggt gacagaaaag cagtgggcca gaggcaccat ggcctcaatg
     2221 tgcttgctgc gctttgttcc atcggaaagc agctcagtct cctgtcagat gggattgatg
     2281 aggaggaatt aattgaggcg gcagacttcc agggcctgta tgctgaggtg aaggcttgct
     2341 cctcagaact ggagagcttg gaaatggagt tgagacaaca gatactggtg aacatcggaa
     2401 agattttaca ggaccagccc agcatggaag ccttagaggc ctcactaggg cagggcctgt
     2461 gcagtggcgg ccaggtggag cctctggacg gcccagctgg ctgcatcctt gagtgtctgg
     2521 tgcttgactc tggagaactg gtgccggaac tcgcagcccc tatcttctac ctgctgggag
     2581 cactggctgt gctgagtgaa acccagcagc agctgctagc taaggctctg gagacaacgg
     2641 tgctgtcaaa gcagctggag ttggtgaagc acgtcttgga acagagcacc ccgtggcagg
     2701 agcagagttc tgtgtccctg cccaccgtgc tccttgggga ctgctgggat gaaaagaatc
     2761 ccacctgggt cttgctagaa gaatgtggcc taaggctgca ggtagaatcc ccccaggtgc
     2821 actgggaacc aacgtctctg atccccacaa gtgcgctcta tgcctccctg ttcctattgt
     2881 caagtctagg ccagaaacct tgtgaacaaa aactcatctc agaagaggat ctgtaggtcg
     2941 acggtatcga taagcttgat atcgaattcc gccccccccc ctaacgttac tggccgaagc
     3001 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     3061 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     3121 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     3181 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     3241 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     3301 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     3361 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     3421 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     3481 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     3541 cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3601 gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc
     3661 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3721 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3781 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3841 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3901 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3961 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     4021 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     4081 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4141 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     4201 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4261 caagtaatga attaattaag aattcgataa aataaaagat tttatttagt ctccagaaaa
     4321 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     4381 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     4441 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     4501 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     4561 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     4621 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     4681 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg
     4741 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     4801 tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga
     4861 agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag
     4921 ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag
     4981 aaagagctgg ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5041 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5101 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     7141 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     7201 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     7261 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     7321 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     7381 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     7441 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     7501 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     7561 gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg
     7621 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     7681 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     7741 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta
     7801 gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat
     7861 caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc
     7921 cagaaaaagg gggga
//
