LOCUS       sequence-361148-        7962 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 7962)
  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 7962)
  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-cat-Caspase-1/4b
FEATURES             Location/Qualifiers
     source          1..7962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..134
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     150..166
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      215..224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             224..253
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1773..2339
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1789..2341
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1933..1950)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2160..2179
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2353..2355,2356..2358,2359..3072)
                     /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(2398..2419)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2659..2678)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3006..3027
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3134..3648
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3810..3826
                     /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(3834..3840,3841..3858,3859..3864))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3879..3900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4017..4034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4188..4776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4268..4287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4947..5738,5739..5807))
                     /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     5570..5589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5808..5912)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5980..5998
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6036..6058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6158..6177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6371..6393
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6521..6540
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6740..7256
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7320..7661
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7728..7962,1..182)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
       61 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
      121 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
      181 ccggaattag atctctcgag atcgatgcgg ccgcgccacc atggaacaaa aactcatctc
      241 agaagaggat ctgatggccg acaaggatct gaagggcaag aggaagcagt tcatcaactc
      301 agtcggcatg gggacggtca acggcttgct ggatgaactc tttgagaaaa acgtgctgaa
      361 ccaggaggag atggagagag taaaatgtga aaacgctacc gttatggaca aggcccgagc
      421 tctgatcgac agcgtcctgc ggaaagggcc acgggcgtgc cagatcttta tctgtcacat
      481 ctgtgaggaa gacacccacc ttgcagagac gctggggctc tcctcaagcc cacaatctgg
      541 aaattctcag aacaccacgg actctgaagt agcgtttcct cctcttccag aggacaaaca
      601 cgacaaaaac ccatttaaga ttttggaatc tgtgggcaaa gagctcctta ctggcgtttt
      661 gcaagacttg gtggagaaag atgtcctgaa attggaggaa gcagaaaaga aaaaatttca
      721 tgatgccaaa cctcgagaca agccccgggt ctttctgaac tcagtgcgtc agaaacccaa
      781 ggaggcaggt caagccattg tccaaacttt ccttaatacg gacaagcatt ctaccagttt
      841 agaagccagc gtgaataaca tgcctgggcc ggctgagcca gaagaatctg tagatgctct
      901 caagctttgt cctcgtgaaa acttcgtgaa actgtgtaaa cagagggctg aagagatcta
      961 cccaataaag gagagaaagg atcgtactcg tctggctctc atcatatgca atacgacgtt
     1021 cgatcatctt tctctcagga agggggctga ccttgacgtt gcagggatga ggaggctgct
     1081 tacagacctt ggctacagtg tgcacataaa agaggaactc actgctaagg acatggaatc
     1141 agagctgagg gcatttgctg cccgtccaga gcacaagtcc tcggacagca cattcctggt
     1201 gttcatgtct catggcatcc tgagtggaat ctgtgggacg aagtacagcg ctgaaggaga
     1261 cccagatgta ttggcttatg acaccatctt ccagattttc aacaaccgca actgccttag
     1321 tctaaaggac aagcccaagg tcatcatcgt ccaggcctgc agaggtgaaa atttggggga
     1381 actgttgatc agtgactctc cagcggcccc aatggacagc acttcacaga tgggtagcag
     1441 cctttcacag gtgggtgaca acctagagga cgacgccatt tacaaggtcc acgtggagaa
     1501 ggacttcatc gctttctgct cttcgacccc acatcatgtg tcttggagag acgtgaacaa
     1561 gggatctctc ttcattacac aactcatcac gtgcttccaa aagtattcgt ggtgctttca
     1621 tctggaggaa gtatttcgga aggtacaaca gtcatttgaa aaaccaaatg ttagagccca
     1681 gatgccgacc attgaacgag tatccatgac aagatatttc tatctcttcc ctggcaattg
     1741 agtcgacggt atcgataagc ttgatatcga attccgcccc cccccctaac gttactggcc
     1801 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     1861 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     1921 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     1981 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2041 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2101 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2161 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2221 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     2281 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2341 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2401 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc
     2461 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2521 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2581 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2641 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2701 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2761 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2821 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc
     2881 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2941 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3001 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3061 ctgtacaagt aatgaattaa ttaagaattc gataaaataa aagattttat ttagtctcca
     3121 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3181 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     3241 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3301 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     3361 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3421 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3481 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     3541 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3601 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     3661 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     3721 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
     3781 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     3841 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3901 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3961 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4021 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4081 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4141 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4201 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4261 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4321 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4381 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4441 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4501 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4561 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4621 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4681 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4741 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4801 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4861 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5821 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5881 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5941 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6001 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6061 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6121 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6181 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6241 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6301 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6361 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     6421 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6481 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6541 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     6601 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     6661 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     6721 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     6781 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     6841 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     6901 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     6961 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     7021 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     7081 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     7141 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
     7201 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
     7261 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
     7321 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
     7381 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     7441 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     7501 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     7561 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     7621 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     7681 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     7741 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     7801 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     7861 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     7921 tcaaggtctt ttcacctggc ccgcatggac acccagacca gg
//
