LOCUS       sequence-361452-        7689 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 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-human-Caspase-1
FEATURES             Location/Qualifiers
     source          1..7689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..104
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     120..136
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      185..194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             194..223
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1470..2036
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1486..2038
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1630..1647)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1857..1876
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2050..2052,2053..2055,2056..2769)
                     /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(2095..2116)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2356..2375)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2703..2724
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2831..3345
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3507..3523
                     /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(3531..3537,3538..3555,3556..3561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3576..3597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3714..3731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3885..4473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3965..3984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4644..5435,5436..5504))
                     /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     5267..5286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5505..5609)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5677..5695
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5733..5755)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5855..5874
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6068..6090
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6218..6237
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6437..6953
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7017..7358
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7425..7689,1..152)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
       61 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
      121 tttatccagc cctcactcct tctctaggcg ccggaattag atctctcgag atcgatgcgg
      181 ccgcgccacc atggaacaaa aactcatctc agaagaggat ctgatggccg acaaggtcct
      241 gaaggagaag agaaagctgt ttatccgttc catgggtgaa ggtacaataa atggcttact
      301 ggatgaatta ttacagacaa gggtgctgaa caaggaagag atggagaaag taaaacgtga
      361 aaatgctaca gttatggata agacccgagc tttgattgac tccgttattc cgaaaggggc
      421 acaggcatgc caaatttgca tcacatacat ttgtgaagaa gacagttacc tggcagggac
      481 gctgggactc tcagcagatc aaacatctgg aaattacctt aatatgcaag actctcaagg
      541 agtactttct tcctttccag ctcctcaggc agtgcaggac aacccagcta tgcccacatc
      601 ctcaggctca gaagggaatg tcaagctttg ctccctagaa gaagctcaaa ggatatggaa
      661 acaaaagtcg gcagagattt atccaataat ggacaagtca agccgcacac gtcttgctct
      721 cattatctgc aatgaagaat ttgacagtat tcctagaaga actggagctg aggttgacat
      781 cacaggcatg acaatgctgc tacaaaatct ggggtacagc gtagatgtga aaaaaaatct
      841 cactgcttcg gacatgacta cagagctgga ggcatttgca caccgcccag agcacaagac
      901 ctctgacagc acgttcctgg tgttcatgtc tcatggtatt cgggaaggca tttgtgggaa
      961 gaaacactct gagcaagtcc cagatatact acaactcaat gcaatcttta acatgttgaa
     1021 taccaagaac tgcccaagtt tgaaggacaa accgaaggtg atcatcatcc aggcctgccg
     1081 tggtgacagc cctggtgtgg tgtggtttaa agattcagta ggagtttctg gaaacctatc
     1141 tttaccaact acagaagagt ttgaggatga tgctattaag aaagcccaca tagagaagga
     1201 ttttatcgct ttctgctctt ccacaccaga taatgtttct tggagacatc ccacaatggg
     1261 ctctgttttt attggaagac tcattgaaca tatgcaagaa tatgcctgtt cctgtgatgt
     1321 ggaggaaatt ttccgcaagg ttcgattttc atttgagcag ccagatggta gagcgcagat
     1381 gcccaccact gaaagagtga ctttgacaag atgtttctac ctcttcccag gacattaagt
     1441 cgacggtatc gataagcttg atatcgaatt ccgccccccc ccctaacgtt actggccgaa
     1501 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1561 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     1621 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     1681 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     1741 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     1801 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     1861 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     1921 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     1981 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2041 gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2101 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat
     2161 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2221 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2281 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2341 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2401 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2461 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2521 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2581 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2641 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2701 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2761 tacaagtaat gaattaatta agaattcgat aaaataaaag attttattta gtctccagaa
     2821 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     2881 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     2941 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3001 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     3061 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     3121 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     3181 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     3241 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     3301 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     3361 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     3421 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     3481 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     3541 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5641 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5701 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5761 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5821 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5881 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5941 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6001 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6061 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga
     6121 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6181 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6241 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat
     6301 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     6361 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc
     6421 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     6481 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     6541 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     6601 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     6661 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     6721 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     6781 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     6841 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc
     6901 cttgggaggg tctcctcaga ttgattgact gcccacctcg ggggtctttc atttggaggt
     6961 tccaccgaga tttggagacc cctgcccagg gaccaccgac ccccccgccg ggaggtaagc
     7021 tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat
     7081 gtttgcgcct gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg
     7141 gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg
     7201 gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat
     7261 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     7321 tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt
     7381 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca
     7441 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt
     7501 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta
     7561 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca
     7621 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg
     7681 aagccttgg
//
