LOCUS       sequence-361167-        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-panda-Caspase-1/4b
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     81..97
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      146..155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             155..184
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1677..2243
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1693..2245
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1837..1854)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2064..2083
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2257..2259,2260..2262,2263..2976)
                     /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(2302..2323)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2563..2582)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2910..2931
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3038..3552
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3714..3730
                     /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(3738..3744,3745..3762,3763..3768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3783..3804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3921..3938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4092..4680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4172..4191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4851..5642,5643..5711))
                     /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     5474..5493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5712..5816)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5884..5902
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5940..5962)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6062..6081
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6275..6297
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6425..6444
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6644..7160
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7224..7565
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7632..7935,1..113)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
       61 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaatta
      121 gatctctcga gatcgatgcg gccgcgccac catggaacaa aaactcatct cagaagagga
      181 tctgatggct gaccagaccc tgagggacaa gcggaagctg tttgtcagct cggtgaacac
      241 ggggaccctc aacggcttgc tggatgagct cttagagaaa agggtgctga accaggagga
      301 gatggagaga gtgcgatatg aaaatgctac agtgatggat aaagcccgag ctctgattga
      361 ctctgtccta cggaaagggt cccaggcatg ccagattttt atttgttata tttgtgatga
      421 ggacgccttc cttgcagaga agatggggct ctcctcaggt acacccttta gaagtcagct
      481 taactcagac tctccagcag caactcttcc ttcaccagaa gacaaacatg acaaaaaccc
      541 atttaagatt ttggaaactg tgggaaaaga actcattact ggtgttttgc atggcttgtt
      601 ggaaaaagat gttctgaaat tggaggaagg agaaaagaaa aagttttacg atgccaaacc
      661 tgaagacaag ccctgggtct ttctgaactc tgtgcggcag aaacggcagg aggcaggtca
      721 agcccttatc caaactttca ttaatacgga caaatattcc acaagtgtaa aagctttcga
      781 ggacatcatg cctgggccac ctgagccaga agaatctacg gatactctca agctttgtcc
      841 tcatgaagaa ttcgtgaaac tgtatacaga gaaggctgga gagatctacc caataaagga
      901 gagacaggat cgtattcgtc tcgctctcat catatgcaat atagaatttg atcatcttcc
      961 tcccaggaat ggggctgaac ttgacattac agggatgaag aatctgctgg aaggccttgg
     1021 ctacagtgtg gatgtgaaac agaaactcac tgctaaggac atggagtcag cgctgcgggc
     1081 atttgctgcc cgcccagagc acgagtcctc agacagcacg ttcctggtgc tcatgtctca
     1141 cggcatcttg agtggaatct gtgggactac gttcagcccg gaaaatccgg atgtgctacc
     1201 ttatgacacc atcttccaga tcttcaacaa ccgcaactgc ttcaaactca gggacaaacc
     1261 caaggtcatc atcatccagg cctgcagagg agaaaatctt ggggaactgt gggtcagtga
     1321 ctctccagca gcctcgactg acagcttttc acatcaacct ctgctcctgg agagtgatgt
     1381 cgtctacaag gtccacgtgg agaaggactt cgttgctttc tgctcctcga ctccacataa
     1441 cgtatcctgg agacacgtca caaagggatc tctcttcatt gcacaactca tcacatgctt
     1501 ccaaaaatat tcttggtgct gtcacctagt agaagtattc cagaaggtac aacaagcttt
     1561 tgaaaaacca aatgtcaaag cccagatgcc caccattgaa cgaatgtcca tgacaaaacc
     1621 tttctatctc ttccctggca actgagtcga cggtatcgat aagcttgata tcgaattccg
     1681 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1741 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     1801 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     1861 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     1921 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     1981 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     2041 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     2101 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     2161 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     2221 tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg
     2281 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2341 agcgtgtctg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2401 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2461 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2521 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2581 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2641 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2701 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc
     2761 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2821 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2881 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2941 gggatcactc tcggcatgga cgagctgtac aagtaatgaa ttaattaaga attcgataaa
     3001 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     3061 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     3121 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
     3181 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     3241 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     3301 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     3361 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     3421 cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     3481 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     3541 ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt
     3601 cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact
     3661 cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat
     3721 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3781 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3841 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3901 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3961 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5761 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5821 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5881 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     5941 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     6001 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     6061 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     6121 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     6181 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     6241 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6301 gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     6361 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     6421 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     6481 ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg
     6541 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     6601 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     6661 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     6721 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     6781 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     6841 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     6901 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     6961 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     7021 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
     7081 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
     7141 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac
     7201 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
     7261 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
     7321 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
     7381 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
     7441 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     7501 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     7561 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     7621 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     7681 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     7741 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     7801 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     7861 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     7921 agccctttgt acacc
//
