LOCUS       sequence-361114-        7926 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 7926)
  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 7926)
  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+human-Caspase-4-catalytic
             domain
FEATURES             Location/Qualifiers
     source          1..7926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..88
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     104..120
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      169..178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             178..207
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1691..2257
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1707..2259
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1851..1868)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2078..2097
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2271..2273,2274..2276,2277..2990)
                     /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(2316..2337)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2577..2596)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2924..2945
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3052..3566
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3728..3744
                     /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(3752..3758,3759..3776,3777..3782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3797..3818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3935..3952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4106..4694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4186..4205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4865..5656,5657..5725))
                     /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     5488..5507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5726..5830)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5898..5916
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5954..5976)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6076..6095
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6289..6311
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6439..6458
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6658..7174
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7238..7579
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7646..7926,1..136)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc
       61 tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac
      121 tccttctcta ggcgccggaa ttagatctct cgagatcgat gcggccgcgc caccatggaa
      181 caaaaactca tctcagaaga ggatctgatg gccgacaagg tcctgaagga caagagaagg
      241 ctgtttgtcc ggtcagtaga catggggacc atcaatggtt tgctggatga actctttgag
      301 aaaagagtgc tgaaccacga ggagatggag cgagtgcggt gtgcacactc tacagttatg
      361 gatcaggccc gagttctgat tgactccgtc cttcggaaag ggccaaatgc atgccagatt
      421 tttatttcta atatttgcaa tgaggacatt cacctggcac agacgctggg gctctcctca
      481 ggttcaccat ctggaaatga tcataccaaa ctagactctc aagtagaagt tccttcttta
      541 ccagaagaca aacacgacaa aaatccattt aagattttgg aaactgtggg caaggaactc
      601 attactggtg ttttgcatgg cttgttggaa aaagatatcc tgaaattgga ggatgcagaa
      661 aagaaaacat tttatgatgc caaacctgaa gacaagccct gggtcattct gaactctgtg
      721 cgacagaaac gccaggaggc aggtcaagcc cttgtccata ctttcattaa tacggacaaa
      781 tattccacca gtgtagaaat ggaggctgga ccacctgagt caggagaatc tacagatgcc
      841 ctcaagcttt gtcctcatga agaattcctg agactatgta aagaaagagc tgaagagatc
      901 tatccaataa aggagagaaa caaccgcaca cgcctggctc tcatcatatg caatacagag
      961 tttgaccatc tgcctccgag gaatggagct gactttgaca tcacagggat gaaggagcta
     1021 cttgagggtc tggactatag tgtagatgta gaagagaatc tgacagccag ggatatggag
     1081 tcagcgctga gggcatttgc taccagacca gagcacaagt cctctgacag cacattcttg
     1141 gtactcatgt ctcatggcat cctggaggga atctgcggaa ctgtgcatga tgagaaaaaa
     1201 ccagatgtgc tgctttatga caccatcttc cagatattca acaaccgcaa ctgcctcagt
     1261 ctgaaggaca aacccaaggt catcattgtc caggcctgca gaggtgcaaa ccgtggggaa
     1321 ctgtgggtca gagactctcc agcatccttg gaagtggcct cttcacagtc atctgagaac
     1381 ctagaggaag atgctgttta caagacccac gtggagaagg acttcattgc tttctgctct
     1441 tcaacgccac acaacgtgtc ctggagagac agcacaatgg gctctatctt catcacacaa
     1501 ctcatcacat gcttccagaa atattcttgg tgctgccacc tagaggaagt atttcggaag
     1561 gtacagcaat catttgaaac tccaagggcc aaagctcaaa tgcccaccat agaacgactg
     1621 tccatgacaa gatatttcta cctctttcct ggcaattgag tcgacggtat cgataagctt
     1681 gatatcgaat tccgcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     1741 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     1801 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     1861 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     1921 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     1981 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     2041 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     2101 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     2161 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     2221 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca
     2281 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2341 acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga
     2401 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2461 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2521 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2581 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2641 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2701 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2761 cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2821 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2881 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2941 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa tgaattaatt
     3001 aagaattcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     3061 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     3121 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     3181 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3241 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3301 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     3361 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     3421 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct
     3481 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     3541 actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt
     3601 ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca
     3661 catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat
     3721 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3781 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3841 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3901 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5881 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     5941 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6001 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6061 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6121 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     6181 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6241 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6301 gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     6361 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6421 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6481 gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga
     6541 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
     6601 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6661 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6721 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     6781 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     6841 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     6901 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     6961 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     7021 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta ttcccaataa
     7081 agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg gtctcctcag
     7141 attgattgac tgcccacctc gggggtcttt catttggagg ttccaccgag atttggagac
     7201 ccctgcccag ggaccaccga cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt
     7261 gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta
     7321 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
     7381 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
     7441 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     7501 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     7561 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
     7621 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     7681 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     7741 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     7801 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     7861 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     7921 ccctcc
//
