LOCUS       sequence-399427-        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_C39A-Myc
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    501..842
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             909..1325
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1255..1277
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1293..1309
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2825..2854
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2889..3455
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2905..3457
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3049..3066)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3276..3295
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3469..3471,3472..3474,3475..4188)
                     /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(3514..3535)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3775..3794)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4122..4143
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4250..4764
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4926..4942
                     /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(4950..4956,4957..4974,4975..4980))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4995..5016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5133..5150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5304..5892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5384..5403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6063..6854,6855..6923))
                     /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     6686..6705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6924..7028)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7096..7114
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7152..7174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7274..7293
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7487..7509
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7637..7656
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7856..7935,1..437)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
       61 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
      121 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
      181 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
      241 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
      301 cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc
      361 atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac
      421 ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac
      481 cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg
      541 tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc
      601 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
      661 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
      721 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
      781 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
      841 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
      901 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
      961 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
     1021 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
     1081 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
     1141 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
     1201 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
     1261 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
     1321 gcgccggaat tagatctctc gagatcgatg cggccgcgcc accatgccat cggcctttga
     1381 gaaagtggtc aagaatgtga tcaaggaggt aagcggcagc agaggcgatc tcattccggt
     1441 ggacagcctg cggaactcca ccagcttcag gccctacgcc cttctgaaca ggaaattttc
     1501 aagctcaagg ttctggaaac cccgttattc atgtgtcaac ctgtcaatca aggacatcct
     1561 ggagcccagt gctccagaac cagaaccgga gtgttttggc tccttcaaag tctctgatgt
     1621 cgtcgatggg aacattcagg gcagagtgat gttgtcaggc atgggagaag ggaaaatttc
     1681 tggtggggct gcagtgtctg acagttccag tgcctccatg aatgtgtgta tactgcgtgt
     1741 gactcagaag acctgggaga ccatgcagca tgaaaggcac cttcagcagc ctgagaacaa
     1801 aatcctgcaa cagcttcgga gtcgtgggga tgacctgttt gtggtgaccg aggtgctgca
     1861 gacaaaggag gaagtgcaga tcactgaggt ccacagccaa gagggctcag gccagtttac
     1921 gctgcctgga gctttatgct tgaagggtga aggcaagggc caccaaagcc ggaagaagat
     1981 ggtgaccatt cctgcaggca gcatcctggc attccgagtg gcccaactgc ttattggctc
     2041 taaatgggat atccttctcg tctcagatga gaaacagagg acctttgagc cctcctcagg
     2101 tgacagaaaa gcagtgggcc agaggcacca tggcctcaat gtgcttgctg cgctttgttc
     2161 catcggaaag cagctcagtc tcctgtcaga tgggattgat gaggaggaat taattgaggc
     2221 ggcagacttc cagggcctgt atgctgaggt gaaggcttgc tcctcagaac tggagagctt
     2281 ggaaatggag ttgagacaac agatactggt gaacatcgga aagattttac aggaccagcc
     2341 cagcatggaa gccttagagg cctcactagg gcagggcctg tgcagtggcg gccaggtgga
     2401 gcctctggac ggcccagctg gctgcatcct tgagtgtctg gtgcttgact ctggagaact
     2461 ggtgccggaa ctcgcagccc ctatcttcta cctgctggga gcactggctg tgctgagtga
     2521 aacccagcag cagctgctag ctaaggctct ggagacaacg gtgctgtcaa agcagctgga
     2581 gttggtgaag cacgtcttgg aacagagcac cccgtggcag gagcagagtt ctgtgtccct
     2641 gcccaccgtg ctccttgggg actgctggga tgaaaagaat cccacctggg tcttgctaga
     2701 agaatgtggc ctaaggctgc aggtagaatc cccccaggtg cactgggaac caacgtctct
     2761 gatccccaca agtgcgctct atgcctccct gttcctattg tcaagtctag gccagaaacc
     2821 ttgtgaacaa aaactcatct cagaagagga tctgtaggtc gacggtatcg ataagcttga
     2881 tatcgaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     2941 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     3001 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     3061 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     3121 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     3181 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     3241 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     3301 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     3361 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     3421 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     3481 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3541 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     3601 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3661 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3721 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3781 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3841 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3901 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     3961 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     4021 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     4081 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     4141 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa
     4201 gaattcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4261 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     4321 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     4381 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     4441 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     4501 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     4561 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     4621 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg
     4681 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     4741 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct
     4801 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca
     4861 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg
     4921 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4981 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5461 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5701 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7021 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7081 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     7141 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     7201 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     7261 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     7321 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     7381 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7441 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7501 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     7561 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     7621 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     7681 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca
     7741 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag
     7801 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     7861 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     7921 aaatacataa ctgag
//
