LOCUS       sequence-361021-        7926 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 7926)
  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 7926)
  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-Mouse-caspase-1/11b
FEATURES             Location/Qualifiers
     source          1..7926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..112
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     128..144
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      193..202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             202..231
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      809..818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1715..2281
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1731..2283
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1875..1892)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2102..2121
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2295..2297,2298..2300,2301..3014)
                     /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(2340..2361)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2601..2620)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2948..2969
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3076..3590
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3752..3768
                     /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(3776..3782,3783..3800,3801..3806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3821..3842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3959..3976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4130..4718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4210..4229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4889..5680,5681..5749))
                     /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     5512..5531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5750..5854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5922..5940
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5978..6000)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6100..6119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6313..6335
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6463..6482
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6682..7198
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7262..7603
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7670..7926,1..160)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc
       61 tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg
      121 atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat ctctcgagat
      181 cgatgcggcc gcgccaccat ggaacaaaaa ctcatctcag aagaggatct gatggctgac
      241 aagatcctga gggcaaagag gaagcaattt atcaactcag tgagtatagg gacaataaat
      301 ggattgttgg atgaactttt agagaagaga gtgctgaatc aggaagaaat ggataaaata
      361 aaacttgcaa acattactgc tatggacaag gcacggaacc tatgtgatca tgtctctaaa
      421 aaagggcccc aggcaagcca aatctttatc acttacattt gtaatgaaga ctgctacctg
      481 gcaggaattc tggagcttca atcagctcca tcagctgaaa catttgttgc tacagaagat
      541 tctaaaggag gacatccttc atcctcagaa aacaaacacc ctgacaaacc acttaaggtg
      601 ttggaacagc tgggcaaaga agtccttacg gagtacctag aaaaattagt acaaagcaat
      661 gtactgaaat taaaggagga agataaacaa aaatttaaca atgctgaacg cagtgacaag
      721 cgttgggttt ttgtagatgc catgaaaaag aaacacagca aagtaggtga aatgcttctc
      781 cagacattct tcagtgtgga cccaggcagc caccatggtg aagctaatct ggaaatggag
      841 gaaccagaag aatcattgaa cactctcaag ctttgttccc ctgaagagtt cacaaggctt
      901 tgcagagaaa agacacaaga aatttaccca ataaaggagg ccaatggccg tacacgaaag
      961 gctcttatca tatgcaatac agagttcaaa catctctcac tgaggtatgg ggctaacttt
     1021 gacatcattg gtatgaaagg ccttcttgaa gacttaggct acgatgtggt ggtgaaagag
     1081 gagcttacag cagagggcat ggagtcagag atgaaagact ttgctgcact ctcagaacac
     1141 cagacatcag acagcacatt cctggtgcta atgtctcatg gcacactgca tggcatttgt
     1201 ggaacaatgc acagtgaaaa aactccagat gtgctacagt atgataccat ctatcagata
     1261 ttcaacaatt gccactgtcc aggtctacga gacaaaccca aagtcatcat tgtgcaggcc
     1321 tgcagaggtg ggaactctgg agaaatgtgg atcagagagt cttcaaaacc ccagttgtgc
     1381 agaggtgtag atctacctag gaatatggaa gctgatgctg tcaagctgag ccacgtggag
     1441 aaggacttca ttgccttcta ctctacaacc ccacatcact tgtcctaccg agacaaaaca
     1501 ggaggctctt acttcatcac tagactcatt tcctgcttcc ggaaacatgc ttgctcttgt
     1561 catctctttg atatattcct gaaggtgcaa caatcatttg aaaaggcaag tattcattcc
     1621 cagatgccca ccattgatcg ggcaaccttg acgagatatt tctacctctt tcctggcaac
     1681 tgagtcgacg gtatcgataa gcttgatatc gaattccgcc ccccccccta acgttactgg
     1741 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     1801 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     1861 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     1921 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     1981 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     2041 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     2101 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     2161 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     2221 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     2281 atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2341 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg
     2401 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2461 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2521 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2581 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2641 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2701 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2761 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggca
     2821 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2881 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2941 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3001 agctgtacaa gtaatgaatt aattaagaat tcgataaaat aaaagatttt atttagtctc
     3061 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     3121 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     3181 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3241 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     3301 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3361 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3421 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     3481 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     3541 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt
     3601 ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact
     3661 caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac
     3721 agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca
     3781 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3841 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3901 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3961 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4021 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4081 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4141 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4201 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4261 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4321 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4381 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4441 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4501 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4561 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4621 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4681 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4741 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4801 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4861 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4921 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4981 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5041 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5101 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5161 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5221 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5281 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5341 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5401 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5461 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5521 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5581 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5641 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5701 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5761 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5821 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5881 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5941 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     6001 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6061 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     6121 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6181 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6241 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     6301 cgccagggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa
     6361 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     6421 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     6481 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     6541 gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca
     6601 acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt
     6661 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     6721 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     6781 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     6841 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     6901 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     6961 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     7021 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     7081 cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggactcgc
     7141 tgatccttgg gagggtctcc tcagattgat tgactgccca cctcgggggt ctttcatttg
     7201 gaggttccac cgagatttgg agacccctgc ccagggacca ccgacccccc cgccgggagg
     7261 taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat
     7321 ctaatgtttg cgcctgcgtc tgtactagtt agctaactag ctctgtatct ggcggacccg
     7381 tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt
     7441 tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc
     7501 aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt
     7561 tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt
     7621 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     7681 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     7741 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     7801 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     7861 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     7921 ctggga
//
