LOCUS       sequence-361023-        7680 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 7680)
  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 7680)
  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-dog-Caspase-1/4a
FEATURES             Location/Qualifiers
     source          1..7680
                     /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"
     CDS             195..224
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1468..2034
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1484..2036
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1628..1645)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1855..1874
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2048..2050,2051..2053,2054..2767)
                     /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(2093..2114)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2354..2373)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2701..2722
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2829..3343
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3505..3521
                     /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(3529..3535,3536..3553,3554..3559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3574..3595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3712..3729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3883..4471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3963..3982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4642..5433,5434..5502))
                     /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     5265..5284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5503..5607)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5675..5693
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5731..5753)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5853..5872
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6066..6088
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6216..6235
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6435..6951
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7015..7356
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7423..7680,1..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 catggaacaa aaactcatct cagaagagga tctggccgac aaggtcctga
      241 aggacaagag aaggctgttt gtccggtcag tagacatggg gaccatcaat ggtttgctgg
      301 atgaactctt tgagaaaaga gtgctgaacc acgaggagat ggagcgagtg cggtgtgcac
      361 actctacagt tatggatcag gcccgagttc tgattgactc cgtccttcgg aaagggccaa
      421 atgcatgcca gatttttatt tctaatattt gcaatgagga cattcacctg gcacagacgc
      481 tggggctctc ctcaggttca ccatctggaa atgatcatac caaactagac tctcaagtag
      541 aagttccttc tttaccagcc ttcgtggaaa acatgcctgg gccaaccatt cctgactcag
      601 aagaatctac agatactctc aagctttgtc ctcctgaaac atttgtgaaa atgtataaag
      661 agaaggctga agagatctac ccaataaagg agagaaagga tcgtactcgt ctggctctca
      721 tcatatgcaa tatagagttt gatcatcttt ctaccaggga tggagctgaa cttgacattg
      781 caggaatgga gagtctgctg gagggcctgg gctacagtgt agttgtgaaa cggaaactca
      841 ctgctaaggg tatggaatca gtgctacggg aatttgctgc ccgcccagag cataagtcct
      901 cagacagcac attcttggtg ttaatgtctc acggcatcct gaatggaatc tgtgggaccg
      961 cacacagcgt ggaaaatcca gatgtactag cttatgacac catcttccag attttcaaca
     1021 accgtcactg cctcaacctc aaggacaaac cgaaggtcat catcatccag gcctgcagag
     1081 gtgaaaatcc tggggaactg tgggtcagcg actctccaaa agcctcgaca gacagctgga
     1141 cacatcaacc tctgatgctc cagagtgatg ccattcacaa ggtccacgtg gagaaggact
     1201 tcattgcttt ctgctcctca accccacata atgtgtcctg gagacatatc acgaagggat
     1261 ctcttttcat tgcacaactc atcacatgct tccaaaaata ttcctggtgc tgtcacctag
     1321 aaggagtatt ccggaaggta caacaatcat ttgaaaaacc agatgtgaaa gcccagatgc
     1381 cgaccattga acgagtatcc atgacaagat atttctatct cttccctggc aattgagtcg
     1441 acggtatcga taagcttgat atcgaattcc gccccccccc ctaacgttac tggccgaagc
     1501 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     1561 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     1621 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     1681 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     1741 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     1801 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     1861 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     1921 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     1981 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     2041 cacaaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2101 gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc
     2161 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2221 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2281 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2341 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2401 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2461 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     2521 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     2581 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2641 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     2701 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2761 caagtaatga attaattaag aattcgataa aataaaagat tttatttagt ctccagaaaa
     2821 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     2881 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     2941 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     3001 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     3061 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     3121 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     3181 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg
     3241 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     3301 tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga
     3361 agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag
     3421 ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag
     3481 aaagagctgg ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5641 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5701 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5761 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5821 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5881 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5941 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6001 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6061 gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg
     6121 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     6181 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     6241 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta
     6301 gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat
     6361 caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc
     6421 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     6481 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     6541 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6601 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     6661 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     6721 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     6781 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     6841 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct
     6901 tgggagggtc tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc
     6961 caccgagatt tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg
     7021 gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt
     7081 ttgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
     7141 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
     7201 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
     7261 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
     7321 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
     7381 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
     7441 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     7501 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     7561 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     7621 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
//
