LOCUS       sequence-361151-        7608 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 7608)
  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 7608)
  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-human-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..139
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     155..171
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      220..229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             229..258
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1424..1990
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1440..1992
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1584..1601)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1811..1830
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2004..2006,2007..2009,2010..2723)
                     /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(2049..2070)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2310..2329)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2657..2678
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2785..3299
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3461..3477
                     /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(3485..3491,3492..3509,3510..3515))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3530..3551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3668..3685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3839..4427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3919..3938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4598..5389,5390..5458))
                     /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     5221..5240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5459..5563)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5631..5649
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5687..5709)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5809..5828
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6022..6044
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6172..6191
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6391..6907
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6971..7312
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7379..7608,1..187)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
       61 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
      121 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
      181 aggcgccgga attagatctc tcgagatcga tgcggccgcg ccaccatgga acaaaaactc
      241 atctcagaag aggatctgat ggcagaaggc aaccacagaa aaaagccact taaggtgttg
      301 gaatccctgg gcaaagattt cctcactggt gttttggata acttggtgga acaaaatgta
      361 ctgaactgga aggaagagga aaaaaagaaa tattacgatg ctaaaactga agacaaagtt
      421 cgggtcatgg cagactctat gcaagagaag caacgtatgg caggacaaat gcttcttcaa
      481 acctttttta acatagacca aatatccccc aataaaaaag ctcatccgaa tatggaggct
      541 ggaccacctg agtcaggaga atctacagat gccctcaagc tttgtcctca tgaagaattc
      601 ctgagactat gtaaagaaag agctgaagag atctatccaa taaaggagag aaacaaccgc
      661 acacgcctgg ctctcatcat atgcaataca gagtttgacc atctgcctcc gaggaatgga
      721 gctgactttg acatcacagg gatgaaggag ctacttgagg gtctggacta tagtgtagat
      781 gtagaagaga atctgacagc cagggatatg gagtcagcgc tgagggcatt tgctaccaga
      841 ccagagcaca agtcctctga cagcacattc ttggtactca tgtctcatgg catcctggag
      901 ggaatctgcg gaactgtgca tgatgagaaa aaaccagatg tgctgcttta tgacaccatc
      961 ttccagatat tcaacaaccg caactgcctc agtctgaagg acaaacccaa ggtcatcatt
     1021 gtccaggcct gcagaggtgc aaaccgtggg gaactgtggg tcagagactc tccagcatcc
     1081 ttggaagtgg cctcttcaca gtcatctgag aacctggagg aagatgctgt ttacaagacc
     1141 cacgtggaga aggacttcat tgctttctgc tcttcaacgc cacacaacgt gtcctggaga
     1201 gacagcacaa tgggctctat cttcatcaca caactcatca catgcttcca gaaatattct
     1261 tggtgctgcc acctagagga agtatttcgg aaggtacagc aatcatttga aactccaagg
     1321 gccaaagctc aaatgcccac catagaacga ctgtccatga caagatattt ctacctcttt
     1381 cctggcaatt gagtcgacgg tatcgataag cttgatatcg aattccgccc ccccccctaa
     1441 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1501 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1561 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     1621 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     1681 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     1741 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     1801 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     1861 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     1921 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     1981 acgatgataa tatggccaca accatggtga gcaagggcga ggagctgttc accggggtgg
     2041 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg
     2101 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2161 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2221 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2281 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2341 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2401 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2461 tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     2521 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2581 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2641 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2701 gcatggacga gctgtacaag taatgaatta attaagaatt cgataaaata aaagatttta
     2761 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     2821 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     2881 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     2941 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     3001 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3061 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3121 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     3181 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     3241 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     3301 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     3361 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     3421 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     3481 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3541 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3601 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3661 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3721 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3781 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3841 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3901 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3961 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4021 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4081 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4141 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4201 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4261 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4321 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4501 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4561 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4681 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4741 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4801 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5041 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5101 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5281 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5521 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5581 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5641 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     5701 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     5761 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     5821 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5881 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     5941 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     6001 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     6061 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6121 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6181 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6241 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     6301 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     6361 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     6421 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     6481 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     6541 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     6601 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     6661 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     6721 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     6781 actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg
     6841 tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc
     6901 tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc
     6961 gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg
     7021 tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg
     7081 gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc
     7141 cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc
     7201 gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     7261 gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg
     7321 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     7381 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     7441 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     7501 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     7561 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccaga
//
