LOCUS       sequence-361116-        7683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of mammalian inflammatory caspase gene in 
            mammalian cells by retroviral transduction .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7683)
  AUTHORS   Devant P, Cao A, Kagan JC
  TITLE     Evolution-inspired redesign of the LPS receptor caspase-4 into an 
            interleukin-1beta converting enzyme.
  JOURNAL   Sci Immunol. 2021 Aug 10;6(62). doi: 10.1126/sciimmunol.abh3567.
  PUBMED    34734155
REFERENCE   2  (bases 1 to 7683)
  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: "Sci 
            Immunol."; date: "2021-08-10"; volume: "6(62). doi"; pages: " 
            10.1126/sciimmunol.abh3567"
COMMENT     Sequence Label: pMSCV-IRES-GFP-mouse-Caspase-1-H340D
FEATURES             Location/Qualifiers
     source          1..7683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..69
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     85..101
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      150..159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             159..188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1429..1995
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1445..1997
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1589..1606)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1816..1835
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2009..2011,2012..2014,2015..2728)
                     /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(2054..2075)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2315..2334)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2662..2683
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2790..3304
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3466..3482
                     /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(3490..3496,3497..3514,3515..3520))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3535..3556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3673..3690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3844..4432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3924..3943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4603..5394,5395..5463))
                     /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     5226..5245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5464..5568)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5636..5654
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5692..5714)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5814..5833
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6027..6049
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6177..6196
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6396..6912
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6976..7317
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7384..7683,1..117)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
       61 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
      121 attagatctc tcgagatcga tgcggccgcg ccaccatgga acaaaaactc atctcagaag
      181 aggatctgat ggctgacaag atcctgaggg caaagaggaa gcaatttatc aactcagtga
      241 gtatagggac aataaatgga ttgttggatg aacttttaga gaagagagtg ctgaatcagg
      301 aagaaatgga taaaataaaa cttgcaaaca ttactgctat ggacaaggca cggaacctat
      361 gtgatcatgt ctctaaaaaa gggccccagg caagccaaat ctttatcact tacatttgta
      421 atgaagactg ctacctggca ggaattctgg agcttcaatc agctccatca gctgaaacat
      481 ttgttgctac agaagattct aaaggaggac atccttcatc ctcagaaaca aaggaagaac
      541 agaacaaaga agatggcaca tttccaggac tgactgggac cctcaagttt tgccctttag
      601 aaaaagccca gaagttatgg aaagaaaatc cttcagagat ttatccaata atgaatacaa
      661 ccactcgtac acgtcttgcc ctcattatct gcaacacaga gtttcaacat ctttctccga
      721 gggttggagc tcaagttgac ctcagagaaa tgaagttgct gctggaggat ctggggtata
      781 ccgtgaaagt gaaagaaaat ctcacagctc tggagatggt gaaagaggtg aaagaatttg
      841 ctgcctgccc agagcacaag acttctgaca gtactttcct tgtattcatg tctcatggta
      901 tccaggaggg aatatgtggg accacatact ctaatgaagt ttcagatatt ttaaaggttg
      961 acacaatctt tcagatgatg aacactttga agtgcccaag cttgaaagac aagcccaagg
     1021 tgatcattat tcaggcatgc cgtggagaga aacaaggagt ggtgttgtta aaagattcag
     1081 taagagactc tgaagaggat ttcttaacgg atgcaatttt tgaagatgat ggcattaaga
     1141 aggcccatat agagaaagat tttattgctt tctgctcttc aacaccagat aatgtgtctt
     1201 ggagagatcc tgtcaggggc tcacttttca ttgagtcact catcaaacac atgaaagaat
     1261 atgcctggtc ttgtgacttg gaggacattt tcagaaaggt tcgattttca tttgaacaac
     1321 cagaatttag gctacagatg cccactgctg atagggtgac cctgacaaaa cgtttctacc
     1381 tcttcccggg acattaagtc gacggtatcg ataagcttga tatcgaattc cgcccccccc
     1441 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     1501 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     1561 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     1621 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     1681 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     1741 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     1801 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     1861 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     1921 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     1981 aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg
     2041 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2101 tggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2161 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2221 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2281 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2341 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2401 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2461 ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     2521 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2581 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2641 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2701 tctcggcatg gacgagctgt acaagtaatg aattaattaa gaattcgata aaataaaaga
     2761 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     2821 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     2881 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     2941 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     3001 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     3061 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     3121 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     3181 tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     3241 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     3301 tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt
     3361 aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat
     3421 agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac
     3481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5581 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5641 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5701 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5761 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     5821 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5881 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5941 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6001 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg
     6061 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     6121 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     6181 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     6241 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
     6301 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     6361 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     6421 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     6481 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     6541 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     6601 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     6661 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     6721 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     6781 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
     6841 aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg
     6901 gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc
     6961 cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt
     7021 gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt
     7081 atctggcgga cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga
     7141 cgtcccaggg actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg
     7201 aatccgaccc cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg
     7261 cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag
     7321 cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat
     7381 tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
     7441 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
     7501 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
     7561 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
     7621 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
     7681 gta
//
