LOCUS       sequence-361166-        7935 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 7935)
  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 7935)
  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-polar-bear-Caspase-1/4b
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..115
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     131..147
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      196..205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             205..234
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1727..2293
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1743..2295
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1887..1904)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2114..2133
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2307..2309,2310..2312,2313..3026)
                     /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(2352..2373)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2613..2632)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2960..2981
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3088..3602
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3764..3780
                     /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(3788..3794,3795..3812,3813..3818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3833..3854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3971..3988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4142..4730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4222..4241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4901..5692,5693..5761))
                     /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     5524..5543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5762..5866)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5934..5952
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5990..6012)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6112..6131
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6325..6347
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6475..6494
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6694..7210
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7274..7615
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7682..7935,1..163)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc
       61 gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc
      121 tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaatta gatctctcga
      181 gatcgatgcg gccgcgccac catggaacaa aaactcatct cagaagagga tctgatggct
      241 gaccagaccc tgagggacaa gcggaagctg tttgtcagct cggtgaacac ggggaccctc
      301 aatggcttgc tggatgagct cttggagaaa agggtgctga accaggagga gatggagaaa
      361 gtgcgatatg aaaacgctac agtgatggat aaagcccggg ctctgatcga ctctgtccta
      421 cggaaagggt cccaggcatg ccagattttt atttgttata tttgtgatga ggactccttc
      481 cttgcagaga agatggggct ctcctcaggt acaccattta gaagtcagct taactcagac
      541 tctccagcag caactcttcc ttcaccagaa gacaaacatg acaaaaaccc atttaagatt
      601 ttggaaactg tgggaaaaga actcattact ggtgttttgc atggcttgtt ggaaaaagat
      661 gttctgaaac tggaggaagg agaaaagaaa aagttttacg atgcgaaacc tgaagacaag
      721 ccccgggtct ttctgaactc tgtgcggcag aaacggcggg aggcaggtca agcccttgtc
      781 caaactttca ttaatacgga caaatattcc acaagtgtaa aagctttcga ggacatcgtg
      841 cctgggccac ctgagccaga agaatctaca gatactctca agctttgtcc tcatgaagaa
      901 ttcgtgaaac tgtatacaga gaaggctgga gagatctacc caataaagga gagacaggat
      961 cgtactcgtc tcgctctcat catatgcaat atagactttg atcatcttcc tcccaggaat
     1021 ggggctgaac ttgacattac agggatgaag aatctgctgg agggccttgg ctacagtgtg
     1081 gatgtgaaac agaaactcac tgctaaggac atggagtcag cgctgcgggc attcgccgcc
     1141 cgcccagagc acgagtcctc agacagcacg ttcctggtgc tcatgtctca cggcatcttg
     1201 agtggaatct gtgggactac gttcagcccg gaaaatccag atgtgctacc ttatgacacc
     1261 gtcttccaga tcttcaacaa ccgcaactgc ttcaaactca gggacaaacc caaggtcatc
     1321 atcatccagg cctgcagagg tgaaaatctt ggggaactgt gggtcagtga ctctccagca
     1381 gcttcgactg acagcttttc acatcaacct ctgctcctgg agagtgacgt cgtctacaag
     1441 gtccacgtgg agaaggactt cgttgctttc tgctcctcga ctccacataa cgtatcctgg
     1501 agacacgtca caaaggggtc tctcttcatt gcacaactca tcgcatgctt ccaaaaatat
     1561 tcttggtgct gtcacctagt agaagtattt cagaaggtac aacaagcttt tgaaaaacca
     1621 aatgtcaaag cccagatgcc caccattgaa cgaatgtcca tgacaaaacc tttctatctc
     1681 ttccctggca actgagtcga cggtatcgat aagcttgata tcgaattccg cccccccccc
     1741 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     1801 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     1861 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     1921 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     1981 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2041 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2101 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2161 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     2221 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     2281 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2341 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtctg
     2401 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2461 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2521 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2581 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2641 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2701 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2761 atatcatggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
     2821 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2881 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2941 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3001 tcggcatgga cgagctgtac aagtaatgaa ttaattaaga attcgataaa ataaaagatt
     3061 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     3121 gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca
     3181 gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag
     3241 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca
     3301 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     3361 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     3421 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg
     3481 cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     3541 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     3601 atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa
     3661 gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag
     3721 ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat ccgctcacaa
     3781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4561 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5881 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5941 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6001 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6061 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6121 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     6181 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6241 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6301 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa
     6361 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     6421 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     6481 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     6541 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     6601 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     6661 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     6721 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     6781 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     6841 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     6901 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     6961 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     7021 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     7081 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa
     7141 tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg
     7201 gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc
     7261 cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt
     7321 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat
     7381 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     7441 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     7501 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     7561 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     7621 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     7681 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     7741 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     7801 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     7861 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     7921 ctacatcgtg acctg
//
