LOCUS       sequence-361169-        7647 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 7647)
  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 7647)
  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-lemur-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..124
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     140..156
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      205..214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             214..243
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1448..2014
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1464..2016
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1608..1625)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1835..1854
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2028..2030,2031..2033,2034..2747)
                     /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(2073..2094)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2334..2353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2681..2702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2809..3323
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3485..3501
                     /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(3509..3515,3516..3533,3534..3539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3554..3575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3692..3709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3863..4451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3943..3962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4622..5413,5414..5482))
                     /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     5245..5264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5483..5587)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5655..5673
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5711..5733)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5833..5852
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6046..6068
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6196..6215
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6415..6931
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6995..7336
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7403..7647,1..172)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
       61 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
      121 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag
      181 atctctcgag atcgatgcgg ccgcgccacc atggaacaaa aactcatctc agaagaggat
      241 ctgatggctg aggacaaaca caaaaacaaa aatccactta agatgttgga agccctgggc
      301 aaagatgtcg ccaatgagtt tctcgatagc ttgatggaaa aaaatgtgct gagagtaaag
      361 gcagaagaca agcaaaattt ttatggtgcc aaaacacaag acaaggcccg agttttgata
      421 tcttctctcc gagacaacca acaggaggcc gttcgattac ttctggaaaa ctaccttcac
      481 ctggaaggaa agcccaccaa tgaagaagct cctccggaaa tcaaggctga gccagctgag
      541 tcagcagaac ctacagatac cctcaagctt tgccctcgta aagacttctt gagactgagt
      601 aaagaacggg ctgatgagat ctaccccata aaggagaaaa aggatcgcac tcgcctggct
      661 ctcatcatat gcaacacaga gtttgaccat cttgagcgga gaaatggggc tgaccatgac
      721 attgtgggga tgaaggggct gcttgagggc ctgggctaca gtgtggatgt aaaagagaaa
      781 ctcacagcca gggatatgga gtctgtgctg cgggaatttg ctgcccgacc agagcacaag
      841 tcctcagaca gcaccttcgt ggtgctcatg tctcatggca tcctggatgg aatctgtgga
      901 acgaagcaca gtgagggaac atgggatgtg ctgccttatg acaccgtctt ccggatactc
      961 aacaaccgca actgccctat tctaaaggac aaacccaagg tcatcattgt ccaggcctgc
     1021 agaggtgcaa accgtgggga agtgtgggtc agagactctc cagaagcctc ggcagccagc
     1081 tcctcacagt tacctgagaa ccctgtggca gacagcttct cacagtcccc tgagaacctg
     1141 gaggaagatg ctgtttgcaa gacccacgtg gagaaggact tcattgcttt ctgctcttcc
     1201 accccacata acatgtcctg gagagacatc acaaagggct ccctcttcat cacacagctc
     1261 atcacatgct tccagaaata ctcttggtgc tgtcacctac aggaagtgtt tcggaaggta
     1321 caacaatcat ttgaaacacc aagtgttcga gcccagatgc ccaccattga acgactgtcc
     1381 atgacaagat gcttctacct ctttcctgga aattgagtcg acggtatcga taagcttgat
     1441 atcgaattcc gccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     1501 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     1561 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     1621 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     1681 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     1741 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     1801 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     1861 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     1921 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     1981 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     2041 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2101 gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2161 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2221 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2281 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2341 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2401 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2461 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggcga
     2521 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2581 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2641 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2701 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga attaattaag
     2761 aattcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     2821 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
     2881 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
     2941 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     3001 ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     3061 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     3121 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     3181 cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc
     3241 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     3301 acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg
     3361 agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat
     3421 actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt
     3481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4261 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5581 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5641 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5701 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5761 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     5821 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     5881 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     5941 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6001 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6061 gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6121 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6181 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6241 ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag
     6301 gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga
     6361 gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     6421 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     6481 aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat
     6541 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     6601 atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag
     6661 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     6721 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     6781 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc ccaataaagc
     6841 ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc tcctcagatt
     6901 gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt tggagacccc
     6961 tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc
     7021 tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta
     7081 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
     7141 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
     7201 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     7261 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     7321 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     7381 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     7441 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     7501 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     7561 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     7621 tggacaccca gaccaggtcc cctacat
//
