LOCUS       sequence-370656-        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
FEATURES             Location/Qualifiers
     source          1..7683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..113
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     129..145
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      194..203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             203..232
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1473..2039
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1489..2041
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1633..1650)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1860..1879
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2053..2055,2056..2058,2059..2772)
                     /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(2098..2119)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2359..2378)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2706..2727
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2834..3348
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3510..3526
                     /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(3534..3540,3541..3558,3559..3564))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3579..3600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3717..3734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3888..4476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3968..3987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4647..5438,5439..5507))
                     /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     5270..5289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5508..5612)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5680..5698
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5736..5758)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5858..5877
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6071..6093
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6221..6240
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6440..6956
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7020..7361
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7428..7683,1..161)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc
       61 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc
      121 gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggaattaga tctctcgaga
      181 tcgatgcggc cgcgccacca tggaacaaaa actcatctca gaagaggatc tgatggctga
      241 caagatcctg agggcaaaga ggaagcaatt tatcaactca gtgagtatag ggacaataaa
      301 tggattgttg gatgaacttt tagagaagag agtgctgaat caggaagaaa tggataaaat
      361 aaaacttgca aacattactg ctatggacaa ggcacggaac ctatgtgatc atgtctctaa
      421 aaaagggccc caggcaagcc aaatctttat cacttacatt tgtaatgaag actgctacct
      481 ggcaggaatt ctggagcttc aatcagctcc atcagctgaa acatttgttg ctacagaaga
      541 ttctaaagga ggacatcctt catcctcaga aacaaaggaa gaacagaaca aagaagatgg
      601 cacatttcca ggactgactg ggaccctcaa gttttgccct ttagaaaaag cccagaagtt
      661 atggaaagaa aatccttcag agatttatcc aataatgaat acaaccactc gtacacgtct
      721 tgccctcatt atctgcaaca cagagtttca acatctttct ccgagggttg gagctcaagt
      781 tgacctcaga gaaatgaagt tgctgctgga ggatctgggg tataccgtga aagtgaaaga
      841 aaatctcaca gctctggaga tggtgaaaga ggtgaaagaa tttgctgcct gcccagagca
      901 caagacttct gacagtactt tccttgtatt catgtctcat ggtatccagg agggaatatg
      961 tgggaccaca tactctaatg aagtttcaga tattttaaag gttgacacaa tctttcagat
     1021 gatgaacact ttgaagtgcc caagcttgaa agacaagccc aaggtgatca ttattcaggc
     1081 atgccgtgga gagaaacaag gagtggtgtt gttaaaagat tcagtaagag actctgaaga
     1141 ggatttctta acggatgcaa tttttgaaga tgatggcatt aagaaggccc atatagagaa
     1201 agattttatt gctttctgct cttcaacacc agataatgtg tcttggagac atcctgtcag
     1261 gggctcactt ttcattgagt cactcatcaa acacatgaaa gaatatgcct ggtcttgtga
     1321 cttggaggac attttcagaa aggttcgatt ttcatttgaa caaccagaat ttaggctaca
     1381 gatgcccact gctgataggg tgaccctgac aaaacgtttc tacctcttcc cgggacatta
     1441 agtcgacggt atcgataagc ttgatatcga attccgcccc cccccctaac gttactggcc
     1501 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     1561 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     1621 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     1681 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     1741 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     1801 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     1861 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     1921 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     1981 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2041 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2101 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc
     2161 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2221 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2281 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2341 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2401 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2461 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2521 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc
     2581 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2641 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2701 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2761 ctgtacaagt aatgaattaa ttaagaattc gataaaataa aagattttat ttagtctcca
     2821 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     2881 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     2941 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3001 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     3061 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3121 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3181 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     3241 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3301 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     3361 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     3421 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
     3481 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     3541 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3601 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3661 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3721 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3781 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3841 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3901 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3961 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4021 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4081 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4141 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4201 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4261 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4321 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4381 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4441 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4501 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4561 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4621 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4681 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4741 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4801 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4861 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4921 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4981 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5041 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5101 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5161 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5221 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5281 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5341 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5401 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5461 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5521 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5641 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5701 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5761 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5821 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     5881 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     5941 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6001 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6061 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     6121 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6181 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6241 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     6301 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     6361 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     6421 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     6481 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     6541 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     6601 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     6661 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     6721 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     6781 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     6841 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
     6901 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
     6961 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
     7021 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
     7081 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     7141 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     7201 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     7261 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     7321 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     7381 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     7441 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     7501 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     7561 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     7621 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     7681 ggg
//
