LOCUS       sequence-361033-        7935 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 7935)
  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 7935)
  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-1/4b
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..137
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     153..169
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             221..250
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1749..2315
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1765..2317
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1909..1926)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2136..2155
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2329..2331,2332..2334,2335..3048)
                     /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(2374..2395)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2635..2654)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2982..3003
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3110..3624
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3786..3802
                     /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(3810..3816,3817..3834,3835..3840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3855..3876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3993..4010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4164..4752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4244..4263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4923..5714,5715..5783))
                     /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     5546..5565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5784..5888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5956..5974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6012..6034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6134..6153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6347..6369
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6497..6516
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6716..7232
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7296..7637
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7704..7935,1..185)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
       61 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
      121 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
      181 gcgccggaat tagatctctc gagatcgatg cggccgcatg gaacaaaaac tcatctcaga
      241 agaggatctg gccgacaagg tcctgaagga caagagaagg ctgtttgtcc ggtcagtaga
      301 catggggacc atcaatggtt tgctggatga actctttgag aaaagagtgc tgaaccacga
      361 ggagatggag cgagtgcggt gtgcacactc tacagttatg gatcaggccc gagttctgat
      421 tgactccgtc cttcggaaag ggccaaatgc atgccagatt tttatttcta atatttgcaa
      481 tgaggacatt cacctggcac agacgctggg gctctcctca ggttcaccat ctggaaatga
      541 tcataccaaa ctagactctc aagtagaagt tccttcttta ccagaagaca aacacgacaa
      601 aaatccattt aagattttgg aaactgtggg caaggaactc attactggtg ttttgcatgg
      661 cttgttggaa aaagatatcc tgaaattgga ggatgcagaa aagaaaacat tttatgatgc
      721 caaacctgaa gacaagccct gggtcattct gaactctgtg cgacagaaac gccaggaggc
      781 aggtcaagcc cttgtccata ctttcattaa tacggacaaa tattccacca gtgtagaagc
      841 cttcgtggaa aacatgcctg ggccaaccat tcctgactca gaagaatcta cagatactct
      901 caagctttgt cctcctgaaa catttgtgaa aatgtataaa gagaaggctg aagagatcta
      961 cccaataaag gagagaaagg atcgtactcg tctggctctc atcatatgca atatagagtt
     1021 tgatcatctt tctaccaggg atggagctga acttgacatt gcaggaatgg agagtctgct
     1081 ggagggcctg ggctacagtg tagttgtgaa acggaaactc actgctaagg gtatggaatc
     1141 agtgctacgg gaatttgctg cccgcccaga gcataagtcc tcagacagca cattcttggt
     1201 gttaatgtct cacggcatcc tgaatggaat ctgtgggacc gcacacagcg tggaaaatcc
     1261 agatgtacta gcttatgaca ccatcttcca gattttcaac aaccgtcact gcctcaacct
     1321 caaggacaaa ccgaaggtca tcatcatcca ggcctgcaga ggtgaaaatc ctggggaact
     1381 gtgggtcagc gactctccaa aagcctcgac agacagctgg acacatcaac ctctgatgct
     1441 ccagagtgat gccattcaca aggtccacgt ggagaaggac ttcattgctt tctgctcctc
     1501 aaccccacat aatgtgtcct ggagacatat cacgaaggga tctcttttca ttgcacaact
     1561 catcacatgc ttccaaaaat attcctggtg ctgtcaccta gaaggagtat tccggaaggt
     1621 acaacaatca tttgaaaaac cagatgtgaa agcccagatg ccgaccattg aacgagtatc
     1681 catgacaaga tatttctatc tcttccctgg caattgagtc gacggtatcg ataagcttga
     1741 tatcgaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     1801 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     1861 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     1921 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     1981 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     2041 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     2101 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     2161 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     2221 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     2281 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     2341 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2401 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     2461 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2521 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2581 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2641 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2701 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2761 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     2821 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2881 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2941 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3001 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa
     3061 gaattcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     3121 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     3181 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     3241 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     3301 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     3361 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     3421 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
     3481 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg
     3541 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     3601 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct
     3661 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca
     3721 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg
     3781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3841 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5881 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5941 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     6001 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     6061 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     6121 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     6181 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     6241 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6301 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6361 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6421 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6481 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6541 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca
     6601 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag
     6661 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     6721 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     6781 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     6841 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     6901 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     6961 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     7021 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     7081 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtatt cccaataaag
     7141 cctcttgctg tttgcatccg aatcgtggac tcgctgatcc ttgggagggt ctcctcagat
     7201 tgattgactg cccacctcgg gggtctttca tttggaggtt ccaccgagat ttggagaccc
     7261 ctgcccaggg accaccgacc cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt
     7321 ctgtctctgt ctttgtgcgt gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact
     7381 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
     7441 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     7501 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     7561 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     7621 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     7681 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     7741 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     7801 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     7861 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     7921 atggacaccc agacc
//
