LOCUS       sequence-361112-        7689 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 7689)
  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 7689)
  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-dog-Caspase-1/4a-triple
            mutant-...
FEATURES             Location/Qualifiers
     source          1..7689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..90
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     106..122
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      171..180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             180..209
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1456..2022
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1472..2024
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1616..1633)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1843..1862
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2036..2038,2039..2041,2042..2755)
                     /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(2081..2102)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2342..2361)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2689..2710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2817..3331
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3493..3509
                     /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(3517..3523,3524..3541,3542..3547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3562..3583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3700..3717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3871..4459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3951..3970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4630..5421,5422..5490))
                     /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     5253..5272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5491..5595)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5663..5681
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5719..5741)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5841..5860
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6054..6076
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6204..6223
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6423..6939
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7003..7344
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7411..7689,1..138)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
       61 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
      121 actccttctc taggcgccgg aattagatct ctcgagatcg atgcggccgc gccaccatgg
      181 aacaaaaact catctcagaa gaggatctga tggccgacaa ggtcctgaag gacaagagaa
      241 ggctgtttgt ccggtcagta gacatgggga ccatcaatgg tttgctggat gaactctttg
      301 agaaaagagt gctgaaccac gaggagatgg agcgagtgcg gtgtgcacac tctacagtta
      361 tggatcaggc ccgagttctg attgactccg tccttcggaa agggccaaat gcatgccaga
      421 tttttatttc taatatttgc aatgaggaca ttcacctggc acagacgctg gggctctcct
      481 caggttcacc atctggaaat gatcatacca aactagactc tcaagtagaa gttccttctt
      541 taccagcctt cgtggaaaac atgcctgggc caaccattcc tgactcagaa gaatctacag
      601 atactctcaa gctttgtcct cctgaaacat ttgtgaaaat gtataaagag aaggctgaag
      661 agatctaccc aataaaggag agaaaggatc gtactcgtct ggctctcatc atatgcaata
      721 tagagtttga tcatctttct accagggatg gagctgaact tgacattgca ggaatggaga
      781 gtctgctgga gggcctgggc tacagtgtag ttgtgaaacg gaaactcact gctaagggta
      841 tggaatcagt gctacgggaa tttgctgccc gcccagagca taagtcctca gacagcacat
      901 tcttggtgtt aatgtctcac ggcatcctga atggaatctg tgggaccgca cacagcgtgg
      961 aaaatccaga tgtactagct tatgacacca tcttccagat tttcaacaac cgtcactgcc
     1021 tcaacctcaa ggacaaaccg aaggtcatca tcatccaggc ctgcagaggt gaaaatcctg
     1081 gggaactgtg ggtcagcgac tctccaaaag cctcgacaga cagctggaca catcaacctc
     1141 tgatgctcca gagtgatgcc attcacaagg tccacgtgga gaaggacttc attgctttct
     1201 gctcctcaac cccacataat gtgtcctgga gagacatcac gatgggatct cttttcattg
     1261 cacaactcat cacatgcttc caaaaatatt cctggtgctg tcacctagaa ggagtattcc
     1321 ggaaggtaca acaatcattt gaaaccccag atgtgaaagc ccagatgccg accattgaac
     1381 gagtatccat gacaagatat ttctatctct tcccaggaaa ttgagtcgac ggtatcgata
     1441 agcttgatat cgaattccgc ccccccccct aacgttactg gccgaagccg cttggaataa
     1501 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     1561 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     1621 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     1681 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     1741 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     1801 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     1861 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     1921 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     1981 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
     2041 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2101 cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca cctacggcaa
     2161 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2221 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2281 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2341 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2401 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2461 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2521 caaggcgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2581 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2641 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2701 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaatgaat
     2761 taattaagaa ttcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     2821 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     2881 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     2941 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     3001 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     3061 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     3121 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     3181 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     3241 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     3301 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     3361 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     3421 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     3481 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3541 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5641 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5701 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5761 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5821 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5881 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5941 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6001 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6061 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6121 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6181 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6241 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     6301 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     6361 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     6421 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     6481 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     6541 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     6601 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     6661 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     6721 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     6781 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtattccc
     6841 aataaagcct cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc
     6901 ctcagattga ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg
     6961 gagacccctg cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt
     7021 ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt
     7081 ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
     7141 tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc
     7201 ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta
     7261 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac
     7321 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg
     7381 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg
     7441 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
     7501 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
     7561 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
     7621 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
     7681 gacccccct
//
