LOCUS       sequence-361179-        7587 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 7587)
  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 7587)
  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-horse-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..100
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     116..132
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      181..190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             190..219
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1364..1930
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1380..1932
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1524..1541)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1751..1770
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1944..1946,1947..1949,1950..2663)
                     /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(1989..2010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2250..2269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2597..2618
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2725..3239
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3401..3417
                     /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(3425..3431,3432..3449,3450..3455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3470..3491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4538..5329,5330..5398))
                     /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     5161..5180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5399..5503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5571..5589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5627..5649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5749..5768
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5962..5984
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6112..6131
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6331..6847
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6911..7252
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7319..7587,1..148)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
       61 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
      121 tccagccctc actccttctc taggcgccgg aattagatct ctcgagatcg atgcggccgc
      181 gccaccatgg aacaaaaact catctcagaa gaggatctga tggctgacag acaagagaga
      241 aatctactta aaactttgga atctgtgggc aaagagctca ttactggcct ttcggttggc
      301 ttggtggaaa aagatgtcct gaaattgaaa gaagaggaaa ggaaaaaatt ttatgaagcc
      361 aaacctggag acaagccctg ggtcttggtg gactctgtgc gagacagacg ccatgaagca
      421 ggtcaaattc tttggcaagc cttgtttaac atagacagaa gtgtcatcag tagaaaagct
      481 cttgaggaaa ttgcggctgg accagctgag tcagcagaac ctacggatac cctcaagctt
      541 tgtcctcatg aagaatacct gagactgtgt aaagaaaggg ctgaagagat ctacccaata
      601 aaggacaaaa aggatcgcac tcgtcgggct ctcatcatat gcaatacaga atttgaccat
      661 ctccctttca ggactggggc tgactttgac ctctcaggga tgaagaggct gctcgaggat
      721 cttggctaca gtgtggatgt gaaagagcaa ctcacagcca cggatatgga gtcagtgctg
      781 cgggaatttg ctgcccacca agatcacagg tcttcagaca gcacattctt agtgttcatg
      841 tctcacggta tcctggatgg aatctgcgga acgacacata gtgatgaaaa accagatgtg
      901 ctaccttacg acaccatctt ccggctattc aacaacagga actgccccag tctaaaggac
      961 aaacctaagg tcatcattgt ccaggcctgc agaggtgcaa atcgtgggga attgtggatc
     1021 cgtgactgtc tagcagcctt ggcacctgag aacttggagg aggacgccgt ttacaagacc
     1081 cacgtggaga aggacttcat tgctttctgc tcctcgaccc cacataatgt gtcctggaga
     1141 gacatcacaa agggatctct cttcattaca caactcatca catgcttcca aagacactct
     1201 tggtgctgtc atgtagagga agtatttcgg aaggtgcaac agtcatttga aaaaccaagt
     1261 attaaaaccc aaatgcccac cactgaacga caatccatga caagatattt ctacctgttt
     1321 cctggcaact gagtcgacgg tatcgataag cttgatatcg aattccgccc ccccccctaa
     1381 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1441 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1501 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     1561 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     1621 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     1681 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     1741 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     1801 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     1861 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     1921 acgatgataa tatggccaca accatggtga gcaagggcga ggagctgttc accggggtgg
     1981 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg
     2041 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2101 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2161 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2221 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2281 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2341 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2401 tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     2461 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2521 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2581 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2641 gcatggacga gctgtacaag taatgaatta attaagaatt cgataaaata aaagatttta
     2701 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     2761 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     2821 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     2881 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     2941 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3001 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3061 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     3121 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     3181 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     3241 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     3301 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     3361 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     3421 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3481 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3541 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4201 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5521 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5581 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     5641 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     5701 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     5761 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5821 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5881 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     5941 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     6001 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6061 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6121 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6181 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     6241 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     6301 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     6361 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     6421 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     6481 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     6541 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     6601 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     6661 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     6721 actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg
     6781 tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc
     6841 tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc
     6901 gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg
     6961 tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg
     7021 gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc
     7081 cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc
     7141 gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     7201 gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg
     7261 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     7321 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     7381 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     7441 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     7501 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     7561 catcgtgacc tgggaagcct tggcttt
//
