LOCUS       sequence-361170-        7641 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 7641)
  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 7641)
  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-sheep-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      161..170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             170..199
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1365..1931
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1381..1933
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1525..1542)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1752..1771
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1945..1947,1948..1950,1951..2664)
                     /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(1990..2011)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2251..2270)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2598..2619
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2726..3240
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3435..3451
                     /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(3459..3465,3466..3483,3484..3489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3504..3525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3642..3659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3813..4401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3893..3912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4572..5363,5364..5432))
                     /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     5195..5214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5433..5537)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5605..5623
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5661..5683)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5783..5802
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5996..6018
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6146..6165
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6365..6881
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6945..7286
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7353..7641,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct ctcgagatcg atgcggccgc gccaccatgg aacaaaaact
      181 catctcagaa gaggatctga tggctgaaga caaacacaac aaaaacccac ttaagacgtt
      241 ggaatctttg ggcaaagaac tcatttctgg ccttttggat gactttgtgg aaaaaaatgt
      301 cctgaaattg gaggaagagg agaagaaaaa aatttatgat gccaaacttc aagacaaagc
      361 ccgggtcttg attgattcta tacgacagaa aaaccaagag gcaggtcaag tctttgttca
      421 aactttccta aacatagaca aaaatcccac caatataaaa gctcctgagg aaactgtggc
      481 tggaccagat gagtcagcag gatctgccgc taccctcaag ctttgccctc atgaagaatt
      541 cctgaaactg tgtaaagaaa gggctggaga gatatatcca ataaaagaaa gaaaggaccg
      601 cactcgtctg gctctcatca tatgcaatac agagtttgat catctgcctc ccaggaatgg
      661 ggctgacctt gacattcttg gaatgaagca gctgcttgag ggtcttggct acactgtgga
      721 agtggaagag aaactcacag ccagagacat ggaatcagtg ctgtggaaat ttgctgcacg
      781 tgaagagcac aaatcctcag acagtacatt cttggtattc atgtctcatg gcatcctgga
      841 tgggatctgt gggactatgc acagagatga ggaaccagat gtattacctt atgacaccat
      901 cttccggaca ttcaacaacc gtaattgcct cagtctaaag gacaaaccta aagtcatcat
      961 tgtccaggcc tgcagaggtg caaatcgtgg ggaattgtgg gtcagtgact ctccagcagc
     1021 cttggcagac agcttttcac agtcacccga gaacctggaa gaggatgctg tttacaagac
     1081 ccacgtggag aaggacttca ttgctttctg ttcctcaact ccacataatg tgtcctggag
     1141 agacatcaaa aaaggttctc tcttcattac aagactcatc acatgcttcc aaaaatatgc
     1201 ttggtgctgt caccttgagg aagtatttag gaaggtacaa caatcatttg aaaaaccaaa
     1261 tgttaaagcc cagatgccca ctgttgaaag actctctatg acaagatatt tctacctctt
     1321 tcctggcaac tgagtcgacg gtatcgataa gcttgatatc gaattccgcc ccccccccta
     1381 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     1441 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1501 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1561 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     1621 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1681 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1741 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1801 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1861 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1921 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     1981 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     2041 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2101 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2161 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2221 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2281 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2341 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2401 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     2461 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2521 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     2581 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2641 ggcatggacg agctgtacaa gtaatgaatt aattaagaat tcgataaaat aaaagatttt
     2701 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     2761 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     2821 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     2881 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     2941 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     3001 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3061 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     3121 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     3181 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     3241 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     3301 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     3361 cattatggac agcgcagaaa gmgctrrrka gwwywttrtr gacakcgcag aaagagctgg
     3421 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3481 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3541 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3601 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3661 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3721 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3781 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3841 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3901 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3961 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4021 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4081 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4141 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4201 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4261 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4321 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4381 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4441 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4501 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4561 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4621 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4681 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4741 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4801 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4861 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4921 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4981 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5041 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5101 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5161 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5221 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5281 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5341 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5401 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5461 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5521 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5581 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5641 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5701 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5761 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     5821 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     5881 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5941 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6001 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     6061 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6121 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6181 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     6241 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     6301 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     6361 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     6421 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     6481 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     6541 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     6601 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     6661 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     6721 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     6781 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     6841 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
     6901 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
     6961 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
     7021 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     7081 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
     7141 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     7201 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     7261 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     7321 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     7381 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     7441 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     7501 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     7561 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     7621 ttgacccccc tccctgggtc a
//
