LOCUS       sequence-361158-        7932 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 7932)
  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 7932)
  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-harbour-seal-Caspase-1/4b
FEATURES             Location/Qualifiers
     source          1..7932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..79
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     95..111
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             169..198
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1688..2254
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1704..2256
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1848..1865)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2075..2094
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2268..2270,2271..2273,2274..2987)
                     /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(2313..2334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2574..2593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2921..2942
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3049..3563
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3725..3741
                     /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(3749..3755,3756..3773,3774..3779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3794..3815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3932..3949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4103..4691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4183..4202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4862..5653,5654..5722))
                     /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     5485..5504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5723..5827)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5895..5913
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5951..5973)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6073..6092
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6286..6308
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6436..6455
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6655..7171
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7235..7576
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7643..7932,1..127)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
       61 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
      121 aggcgccgga attagatctc tcgagatcga tgcggccgcg ccaccatgga acaaaaactc
      181 atctcagaag aggatctgat ggctgacaag accctgaagg acaagcggaa gctgtttgtc
      241 agctcagtga acacagggac cctcaacggc ttgctggatg agctcttaga gaaaagagtg
      301 ctgaaccagg aggagatgga gagagtgcgg tatgaaaatg ctacggtgat ggataaggcc
      361 cgagctctga ttgactctgt tgttcggaaa gggtcacagg catgcgatat ttttatttgt
      421 tatattcgtg aagaggactc cttccttgca gagaagatgg ggctttcctc aggtgcacca
      481 tctggaagtc ggcttaactc agactctcaa gcagcaattc ttccttcacc agaagacaaa
      541 catgacaaaa acccatttaa gattttggaa actgtgggca aagaactcat tactggtgtt
      601 ttgcatggct tgttggaaaa agatgttctg aaattggagg aagcagaaaa gaaaaaattt
      661 tatgatgcca aacctgaaga caagccctgg gtctttctgg actctgtgcg acagaaacgg
      721 caggaggcag gtcaagccct tgtccaaact ttcattaaaa cagacaaata ttccaccagt
      781 gtaaaaggtc ccgaggacat gcctgggcaa cctgagccag aagaatctac agatactctc
      841 aagctttgtc ctcatgaaga attcatgaaa ctgtataatg agaaggctgc agagatctac
      901 ccaataaagg agaaaaagga tcgtactcgt ctcgctctca tcatatgcaa tatagagttc
      961 gatcatcttc ctcccaggga tggggccaaa cttgacatcg cagggatgag gagtctactg
     1021 gagggccttg gctacagtgt ggatgtgaaa gaggacctca ctgctaagga tatggagtca
     1081 gcgctgaggg cattcgccgc ccgcccagag cacaagtgct cagacagcac gttcctggtg
     1141 ctcatgtctc acggaatcct gagtggaatc tgcgggactg ccttcagccc ggaaaacccg
     1201 gatgtgctac cttatgacac catcttccag atcttcaaca accgtaactg cctcaacctc
     1261 agggacaaac ccaaggtcat catcatccag gcctgcagag gtgaaagtct tggggaagtg
     1321 tgggtcagtg actcaccagc ggcctcaacc aacagcttct cacatcgacc tctgctcctg
     1381 gagagtgatg ccgtccagaa ggtccacatc gagaaggact tcgttgcttt ctgctcctcg
     1441 accccacata acgtatcctg gagacacgtc acaaagggat ctctcttcat tgtacaactc
     1501 atcacatgct tccaaaaata ttcttggagc tatcacctca tagaagtatt tcagaaggta
     1561 caaaaatcat ttgaaaaacc aaatgttaaa gcccagatgc ccaccattga acgaatgtcc
     1621 atgacaagac ctttctatct cttccctggc aattgagtcg acggtatcga taagcttgat
     1681 atcgaattcc gccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     1741 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     1801 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     1861 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     1921 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     1981 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     2041 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     2101 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     2161 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     2221 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     2281 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2341 gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2401 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2461 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2521 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2581 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2641 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2701 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggcga
     2761 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2821 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2881 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2941 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga attaattaag
     3001 aattcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     3061 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     3121 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     3181 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     3241 ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     3301 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     3361 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     3421 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc
     3481 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     3541 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg
     3601 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat
     3661 actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt
     3721 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3781 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3841 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3901 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4861 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4921 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4981 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5041 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5101 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5161 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5221 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5281 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5341 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5401 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5461 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5521 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5581 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5641 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5701 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5761 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5821 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5881 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5941 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     6001 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     6061 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6121 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     6181 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6241 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6301 gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6361 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6421 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6481 ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag
     6541 gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga
     6601 gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     6661 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     6721 aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat
     6781 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     6841 atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag
     6901 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     6961 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     7021 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc ccaataaagc
     7081 ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt
     7141 gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc
     7201 tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc
     7261 tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta
     7321 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
     7381 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
     7441 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     7501 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     7561 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     7621 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     7681 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     7741 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     7801 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     7861 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     7921 tccctgggtc aa
//
