LOCUS       sequence-370659-        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-1/4a
FEATURES             Location/Qualifiers
     source          1..7689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..96
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     112..128
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      177..186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             186..215
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1462..2028
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1478..2030
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1622..1639)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1849..1868
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2042..2044,2045..2047,2048..2761)
                     /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(2087..2108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2348..2367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2695..2716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2823..3337
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3499..3515
                     /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(3523..3529,3530..3547,3548..3553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3568..3589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3706..3723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3877..4465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3957..3976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4636..5427,5428..5496))
                     /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     5259..5278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5497..5601)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5669..5687
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5725..5747)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5847..5866
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6060..6082
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6210..6229
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6429..6945
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7009..7350
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7417..7689,1..144)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
       61 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
      121 gccctcactc cttctctagg cgccggaatt agatctctcg agatcgatgc ggccgcgcca
      181 ccatggaaca aaaactcatc tcagaagagg atctgatggc cgacaaggtc ctgaaggaga
      241 agagaaagct gtttatccgt tccatgggtg aaggtacaat aaatggctta ctggatgaat
      301 tattacagac aagggtgctg aacaaggaag agatggagaa agtaaaacgt gaaaatgcta
      361 cagttatgga taagacccga gctttgattg actccgttat tccgaaaggg gcacaggcat
      421 gccaaatttg catcacatac atttgtgaag aagacagtta cctggcaggg acgctgggac
      481 tctcagcaga tcaaacatct ggaaattacc ttaatatgca agactctcaa ggagtacttt
      541 cttcctttcc aaaaaaagct catccgaata tggaggctgg accacctgag tcaggagaat
      601 ctacagatgc cctcaagctt tgtcctcatg aagaattcct gagactatgt aaagaaagag
      661 ctgaagagat ctatccaata aaggagagaa acaaccgcac acgcctggct ctcatcatat
      721 gcaatacaga gtttgaccat ctgcctccga ggaatggagc tgactttgac atcacaggga
      781 tgaaggagct acttgagggt ctggactata gtgtagatgt agaagagaat ctgacagcca
      841 gggatatgga gtcagcgctg agggcatttg ctaccagacc agagcacaag tcctctgaca
      901 gcacattctt ggtactcatg tctcatggca tcctggaggg aatctgcgga actgtgcatg
      961 atgagaaaaa accagatgtg ctgctttatg acaccatctt ccagatattc aacaaccgca
     1021 actgcctcag tctgaaggac aaacccaagg tcatcattgt ccaggcctgc agaggtgcaa
     1081 accgtgggga actgtgggtc agagactctc cagcatcctt ggaagtggcc tcttcacagt
     1141 catctgagaa cctagaggaa gatgctgttt acaagaccca cgtggagaag gacttcattg
     1201 ctttctgctc ttcaacgcca cacaacgtgt cctggagaga cagcacaatg ggctctatct
     1261 tcatcacaca actcatcaca tgcttccaga aatattcttg gtgctgccac ctagaggaag
     1321 tatttcggaa ggtacagcaa tcatttgaaa ctccaagggc caaagctcaa atgcccacca
     1381 tagaacgact gtccatgaca agatatttct acctctttcc tggcaattga gtcgacggta
     1441 tcgataagct tgatatcgaa ttccgccccc ccccctaacg ttactggccg aagccgcttg
     1501 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     1561 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     1621 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     1681 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     1741 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     1801 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     1861 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     1921 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     1981 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     2041 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2101 cggcgacgta aacggccaca agttcagcgt gtctggcgag ggcgagggcg atgccaccta
     2161 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2221 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2281 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2341 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2401 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2461 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2521 cggcatcaag gcgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2581 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2641 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2701 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2761 atgaattaat taagaattcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     2821 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     2881 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     2941 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3001 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     3061 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3121 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3181 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     3241 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3301 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     3361 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     3421 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     3481 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5641 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5701 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5761 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5821 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5881 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5941 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6001 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6061 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     6121 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6181 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6241 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     6301 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     6361 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     6421 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     6481 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
     6541 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6601 agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca
     6661 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     6721 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     6781 cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt
     6841 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
     6901 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
     6961 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
     7021 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
     7081 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     7141 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
     7201 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
     7261 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     7321 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
     7381 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
     7441 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
     7501 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
     7561 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
     7621 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
     7681 gcttttgac
//
