LOCUS       sequence-370661-        7590 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 7590)
  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 7590)
  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-rabbit-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..95
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     111..127
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      176..185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             185..214
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1362..1928
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1378..1930
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1522..1539)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1749..1768
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1942..1944,1945..1947,1948..2661)
                     /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(1987..2008)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2248..2267)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2595..2616
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2723..3237
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3399..3415
                     /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(3423..3429,3430..3447,3448..3453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3468..3489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3606..3623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3777..4365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3857..3876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4536..5327,5328..5396))
                     /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     5159..5178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5397..5501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5569..5587
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5625..5647)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5747..5766
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5960..5982
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6110..6129
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6329..6845
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6909..7250
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7317..7590,1..143)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
       61 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
      121 ccctcactcc ttctctaggc gccggaatta gatctctcga gatcgatgcg gccgcgccac
      181 catggaacaa aaactcatct cagaagagga tctgatggct gaatacaagg atgtacttaa
      241 ggagatggaa tccgcgggca aagctattgc aggtagcttt atggataatt tggtgaaaaa
      301 aaaagtgata aaattggaga aagcgaagag ggaaaagttt tatgatgcca aaactcacaa
      361 aaaggcccaa atcttggtgg actctcttcg agagaagtgc catgaagaag atcaaaagct
      421 tactgaaacc tttcttaaca ttgaccaaaa ttctactggt atgcaagaaa tcaaggctgg
      481 accagaggat tcagcagaat ctacagatgt tctcaagctt tgtccatctg aaaaattcct
      541 gagactctgt aaagaaagag ctggagagat ctacccaata aaggacaaaa aggatcgcac
      601 tcgcctggct ctcatcatat gcaacataga gtttcagcat cttcctctga ggaatggggc
      661 tcagtttgac atctcaggga tgaagaggct gcttgaggac ctgggttaca gtgtggttgt
      721 gaaagtgaaa ctcacagcca aggctatgga gtcagtgctg caggagttcg ctgctcggcc
      781 agagcacaat tcttcagata gcacatttgt ggtgctcatg tctcatggca tcctggatgg
      841 aatctgtggc attatgcata gtgacaaaac acaagatgtg ctaccttatg acactgtatt
      901 ccggacactc aacaaccgca actgcatcag tctgagggac aaacccaagg tcatcattgt
      961 acaggcctgc aggggtgcaa atcatggaga agtgtggttt agtgactctc caatggcctt
     1021 ggcaaacagc tcttcacagt tgcctgagaa tctgcaggat gatggcattt acaagaccca
     1081 tgtggagaag gatttcattg ctttctgctc ttcaacacca cataatctgt cctggagaga
     1141 catcacaaag ggctctctgt tcatcacaca actcatctcc tgcttccaaa agtattcctg
     1201 gtgctgtcat ctagaggaag tatttcggaa ggtccaacaa tcatttgaaa aaccaagtgt
     1261 gaaagcccag atgcccacca tcgagcgact gtctatgaca agatacttct ttctctttcc
     1321 tggtaactga gtcgacggta tcgataagct tgatatcgaa ttccgccccc ccccctaacg
     1381 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1441 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1501 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1561 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1621 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1681 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1741 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1801 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     1861 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1921 gatgataata tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1981 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtctggcgag
     2041 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2101 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2161 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2221 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2281 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2341 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2401 atggccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacatcgag
     2461 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2521 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2581 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2641 atggacgagc tgtacaagta atgaattaat taagaattcg ataaaataaa agattttatt
     2701 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     2761 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     2821 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     2881 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     2941 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     3001 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     3061 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     3121 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     3181 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg
     3241 taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat
     3301 cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat
     3361 tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca
     3421 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3961 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4201 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4381 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4741 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4801 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4861 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4921 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4981 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5041 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5101 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5161 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5221 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5281 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5341 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5521 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5581 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5641 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5701 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     5761 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5821 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5881 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     5941 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg
     6001 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6061 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6121 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6181 cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt
     6241 tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga
     6301 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     6361 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     6421 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     6481 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     6541 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     6601 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     6661 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     6721 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
     6781 gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct cgggggtctt
     6841 tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg acccccccgc
     6901 cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg
     6961 ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     7021 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     7081 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     7141 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     7201 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     7261 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     7321 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     7381 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     7441 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     7501 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     7561 tcgtgacctg ggaagccttg gcttttgacc
//
