LOCUS       sequence-361185-        7626 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 7626)
  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 7626)
  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-Hybrid-caspase_mouse-caspase-11-CARD+canine-Caspase-1
            /4-catalytic domain
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..111
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     127..143
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      192..201
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             201..230
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      478..487
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1414..1980
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1430..1982
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1574..1591)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1801..1820
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1994..1996,1997..1999,2000..2713)
                     /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(2039..2060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2300..2319)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2647..2668
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2775..3289
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3451..3467
                     /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(3475..3481,3482..3499,3500..3505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3520..3541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3658..3675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3829..4417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3909..3928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4588..5379,5380..5448))
                     /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     5211..5230
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5449..5553)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5621..5639
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5677..5699)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5799..5818
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6012..6034
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6162..6181
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6381..6897
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6961..7302
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7369..7626,1..159)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
       61 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
      121 tcctcccttt atccagccct cactccttct ctaggcgccg gaattagatc tctcgagatc
      181 gatgcggccg cgccaccatg gaacaaaaac tcatctcaga agaggatctg atggctgaaa
      241 acaaacaccc tgacaaacca cttaaggtgt tggaacagct gggcaaagaa gtccttacgg
      301 agtacctaga aaaattagta caaagcaatg tactgaaatt aaaggaggaa gataaacaaa
      361 aatttaacaa tgctgaacgc agtgacaagc gttgggtttt tgtagatgcc atgaaaaaga
      421 aacacagcaa agtaggtgaa atgcttctcc agacattctt cagtgtggac ccaggcagcc
      481 accatggtga agctaatctg gaagccttcg tggaaaacat gcctgggcca accattcctg
      541 actcagaaga atctacagat actctcaagc tttgtcctcc tgaaacattt gtgaaaatgt
      601 ataaagagaa ggctgaagag atctacccaa taaaggagag aaaggatcgt actcgtctgg
      661 ctctcatcat atgcaatata gagtttgatc atctttctac cagggatgga gctgaacttg
      721 acattgcagg aatggagagt ctgctggagg gcctgggcta cagtgtagtt gtgaaacgga
      781 aactcactgc taagggtatg gaatcagtgc tacgggaatt tgctgcccgc ccagagcata
      841 agtcctcaga cagcacattc ttggtgttaa tgtctcacgg catcctgaat ggaatctgtg
      901 ggaccgcaca cagcgtggaa aatccagatg tactagctta tgacaccatc ttccagattt
      961 tcaacaaccg tcactgcctc aacctcaagg acaaaccgaa ggtcatcatc atccaggcct
     1021 gcagaggtga aaatcctggg gaactgtggg tcagcgactc tccaaaagcc tcgacagaca
     1081 gctggacaca tcaacctctg atgctccaga gtgatgccat tcacaaggtc cacgtggaga
     1141 aggacttcat tgctttctgc tcctcaaccc cacataatgt gtcctggaga catatcacga
     1201 agggatctct tttcattgca caactcatca catgcttcca aaaatattcc tggtgctgtc
     1261 acctagaagg agtattccgg aaggtacaac aatcatttga aaaaccagat gtgaaagccc
     1321 agatgccgac cattgaacga gtatccatga caagatattt ctatctcttc ccaggacatt
     1381 aagtcgacgg tatcgataag cttgatatcg aattccgccc ccccccctaa cgttactggc
     1441 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1501 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1561 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1621 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     1681 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1741 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     1801 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     1861 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     1921 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     1981 tatggccaca accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2041 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg
     2101 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2161 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2221 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2281 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2341 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2401 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2461 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag
     2521 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2581 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2641 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2701 gctgtacaag taatgaatta attaagaatt cgataaaata aaagatttta tttagtctcc
     2761 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     2821 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     2881 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     2941 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     3001 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     3061 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     3121 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     3181 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     3241 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt
     3301 tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc
     3361 aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca
     3421 gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat
     3481 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3541 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3601 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3661 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3721 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3781 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3841 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3901 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3961 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4021 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4081 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4141 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4201 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4261 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4321 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4381 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4441 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4501 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4561 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4621 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4681 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4741 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4801 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4861 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4921 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4981 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5041 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5101 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5161 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5221 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5281 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5341 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5401 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5461 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5521 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5581 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5641 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5701 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5761 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5821 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5881 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5941 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6001 gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag
     6061 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     6121 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     6181 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     6241 cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa
     6301 caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt
     6361 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     6421 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
     6481 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     6541 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     6601 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     6661 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     6721 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     6781 ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct
     6841 gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg
     6901 aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt
     6961 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc
     7021 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt
     7081 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
     7141 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
     7201 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     7261 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
     7321 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
     7381 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     7441 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     7501 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     7561 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     7621 tgggaa
//
