LOCUS       sequence-361184-        7626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 7626)
  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 7626)
  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-Hybrid-caspase_human-caspase-4-CARD+canine-Caspase-1/
            4-catalytic domain
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     81..103
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     119..135
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      184..193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             193..222
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1406..1972
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1422..1974
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1566..1583)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1793..1812
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1986..1988,1989..1991,1992..2705)
                     /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(2031..2052)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2292..2311)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2639..2660
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2767..3281
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3443..3459
                     /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(3467..3473,3474..3491,3492..3497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3512..3533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3650..3667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3821..4409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3901..3920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4580..5371,5372..5440))
                     /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     5203..5222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5441..5545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5613..5631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5669..5691)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5791..5810
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6004..6026
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6154..6173
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6373..6889
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6953..7294
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7361..7626,1..151)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
       61 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
      121 ttatccagcc ctcactcctt ctctaggcgc cggaattaga tctctcgaga tcgatgcggc
      181 cgcgccacca tggaacaaaa actcatctca gaagaggatc tgatggcaga aggcaaccac
      241 agaaaaaagc cacttaaggt gttggaatcc ctgggcaaag atttcctcac tggtgttttg
      301 gataacttgg tggaacaaaa tgtactgaac tggaaggaag aggaaaaaaa gaaatattac
      361 gatgctaaaa ctgaagacaa agttcgggtc atggcagact ctatgcaaga gaagcaacgt
      421 atggcaggac aaatgcttct tcaaaccttt tttaacatag accaaatatc ccccaataaa
      481 aaagctcatc cgaatgcctt cgtggaaaac atgcctgggc caaccattcc tgactcagaa
      541 gaatctacag atactctcaa gctttgtcct cctgaaacat ttgtgaaaat gtataaagag
      601 aaggctgaag agatctaccc aataaaggag agaaaggatc gtactcgtct ggctctcatc
      661 atatgcaata tagagtttga tcatctttct accagggatg gagctgaact tgacattgca
      721 ggaatggaga gtctgctgga gggcctgggc tacagtgtag ttgtgaaacg gaaactcact
      781 gctaagggta tggaatcagt gctacgggaa tttgctgccc gcccagagca taagtcctca
      841 gacagcacat tcttggtgtt aatgtctcac ggcatcctga atggaatctg tgggaccgca
      901 cacagcgtgg aaaatccaga tgtactagct tatgacacca tcttccagat tttcaacaac
      961 cgtcactgcc tcaacctcaa ggacaaaccg aaggtcatca tcatccaggc ctgcagaggt
     1021 gaaaatcctg gggaactgtg ggtcagcgac tctccaaaag cctcgacaga cagctggaca
     1081 catcaacctc tgatgctcca gagtgatgcc attcacaagg tccacgtgga gaaggacttc
     1141 attgctttct gctcctcaac cccacataat gtgtcctgga gacatatcac gaagggatct
     1201 cttttcattg cacaactcat cacatgcttc caaaaatatt cctggtgctg tcacctagaa
     1261 ggagtattcc ggaaggtaca acaatcattt gaaaaaccag atgtgaaagc ccagatgccg
     1321 accattgaac gagtatccat gacaagatat ttctatctct tcccaggaca ttaagtcgac
     1381 ggtatcgata agcttgatat cgaattccgc ccccccccct aacgttactg gccgaagccg
     1441 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     1501 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     1561 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     1621 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     1681 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     1741 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     1801 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     1861 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     1921 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     1981 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2041 tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca
     2101 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2161 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2221 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2281 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2341 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2401 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2461 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2521 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2581 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2641 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2701 agtaatgaat taattaagaa ttcgataaaa taaaagattt tatttagtct ccagaaaaag
     2761 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     2821 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     2881 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     2941 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     3001 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     3061 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     3121 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta
     3181 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     3241 tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag
     3301 ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc
     3361 actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa
     3421 agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3601 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3661 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3721 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3781 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3841 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3901 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3961 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4021 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4081 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4141 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4201 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4261 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4321 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4441 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5581 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5641 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5701 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5761 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5821 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     5881 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5941 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6001 tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc
     6061 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     6121 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     6181 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt
     6241 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     6301 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     6361 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     6421 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     6481 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     6541 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     6601 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     6661 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     6721 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     6781 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg ctgatccttg
     6841 ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt ggaggttcca
     6901 ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag gtaagctggc
     6961 cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt
     7021 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     7081 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     7141 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     7201 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     7261 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     7321 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     7381 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     7441 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     7501 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     7561 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     7621 cttggc
//
