LOCUS       sequence-361120-        7904 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 7904)
  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 7904)
  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-Hybrid-caspase_canine-CARDs+mouse-Caspase-11-catalyti
            c domain
FEATURES             Location/Qualifiers
     source          1..7904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..84
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     100..116
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      165..174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             174..203
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1675..2241
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1691..2243
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1835..1852)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2062..2081
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2255..2257,2258..2260,2261..2974)
                     /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(2300..2321)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2561..2580)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2908..2929
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3036..3550
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3712..3728
                     /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(3736..3742,3743..3760,3761..3766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3781..3802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3919..3936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4090..4678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4170..4189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4849..5640,5641..5709))
                     /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     5472..5491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5710..5814)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5882..5900
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5938..5960)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6060..6079
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6263..6285
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6413..6432
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6632..7148
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7212..7553
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7620..7904,1..132)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
       61 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
      121 tctctaggcg ccggaattag atctctcgag atcgatgcgg ccgcgccacc atggaacaaa
      181 aactcatctc agaagaggat ctgatggccg acaaggtcct gaaggacaag agaaggctgt
      241 ttgtccggtc agtagacatg gggaccatca atggtttgct ggatgaactc tttgagaaaa
      301 gagtgctgaa ccacgaggag atggagcgag tgcggtgtgc acactctaca gttatggatc
      361 aggcccgagt tctgattgac tccgtccttc ggaaagggcc aaatgcatgc cagattttta
      421 tttctaatat ttgcaatgag gacattcacc tggcacagac gctggggctc tcctcaggtt
      481 caccatctgg aaatgatcat accaaactag actctcaagt agaagttcct tctttaccag
      541 aagacaaaca cgacaaaaat ccatttaaga ttttggaaac tgtgggcaag gaactcatta
      601 ctggtgtttt gcatggcttg ttggaaaaag atatcctgaa attggaggat gcagaaaaga
      661 aaacatttta tgatgccaaa cctgaagaca agccctgggt cattctgaac tctgtgcgac
      721 agaaacgcca ggaggcaggt caagcccttg tccatacttt cattaatacg gacaaatatt
      781 ccaccagtgt agaaatggag gaaccagaag aatcattgaa cactctcaag ctttgttccc
      841 ctgaagagtt cacaaggctt tgcagagaaa agacacaaga aatttaccca ataaaggagg
      901 ccaatggccg tacacgaaag gctcttatca tatgcaatac agagttcaaa catctctcac
      961 tgaggtatgg ggctaacttt gacatcattg gtatgaaagg ccttcttgaa gacttaggct
     1021 acgatgtggt ggtgaaagag gagcttacag cagagggcat ggagtcagag atgaaagact
     1081 ttgctgcact ctcagaacac cagacatcag acagcacatt cctggtgcta atgtctcatg
     1141 gcacactgca tggcatttgt ggaacaatgc acagtgaaaa aactccagat gtgctacagt
     1201 atgataccat ctatcagata ttcaacaatt gccactgtcc aggtctacga gacaaaccca
     1261 aagtcatcat tgtgcaggcc tgcagaggtg ggaactctgg agaaatgtgg atcagagagt
     1321 cttcaaaacc ccagttgtgc agaggtgtag atctacctag gaatatggaa gctgatgctg
     1381 tcaagctgag ccacgtggag aaggacttca ttgccttcta ctctacaacc ccacatcact
     1441 tgtcctaccg agacaaaaca ggaggctctt acttcatcac tagactcatt tcctgcttcc
     1501 ggaaacatgc ttgctcttgt catctctttg atatattcct gaaggtgcaa caatcatttg
     1561 aaaaggcaag tattcattcc cagatgccca ccattgatcg ggcaaccttg acgagatatt
     1621 tctacctctt tcctggcaac tgagtcgacg gtatcgataa gcttgatatc gaattccgcc
     1681 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     1741 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     1801 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     1861 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     1921 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     1981 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2041 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2101 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     2161 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2221 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     2281 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2341 cgtgtctggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2401 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2461 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2521 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2581 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2641 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2701 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg
     2761 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2821 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2881 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2941 gatcactctc ggcatggacg agctgtacaa gtaatgaatt aattaagaat tcgataaaat
     3001 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     3061 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     3121 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     3181 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     3241 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     3301 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     3361 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     3421 atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     3481 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     3541 ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg
     3601 aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc
     3661 tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc
     3721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5821 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5881 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5941 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6001 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6061 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6121 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6241 aaggcgatta agttggggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg
     6301 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6361 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6421 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6481 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     6541 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     6601 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     6661 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     6721 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
     6781 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
     6841 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     6901 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     6961 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     7021 gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc
     7081 gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca cctcgggggt
     7141 ctttcatttg gaggttccac cgagatttgg agacccctgc ccagggacca ccgacccccc
     7201 cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt
     7261 gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt agctaactag ctctgtatct
     7321 ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc
     7381 ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc
     7441 cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
     7501 cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct
     7561 gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg
     7621 gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga
     7681 tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag
     7741 aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca
     7801 cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct
     7861 acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgg
//
