LOCUS       sequence-361154-        7596 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 7596)
  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 7596)
  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-mouse-Caspase-11
FEATURES             Location/Qualifiers
     source          1..7596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..118
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     134..150
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      199..208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             208..237
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      485..494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1391..1957
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1407..1959
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1551..1568)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1778..1797
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1971..1973,1974..1976,1977..2690)
                     /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(2016..2037)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2277..2296)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2624..2645
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2752..3266
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3428..3444
                     /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(3452..3458,3459..3476,3477..3482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3497..3518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3635..3652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3806..4394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3886..3905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4565..5356,5357..5425))
                     /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     5188..5207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5426..5530)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5598..5616
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5654..5676)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5776..5795
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5989..6011
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6139..6158
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6358..6874
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6938..7279
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7346..7596,1..166)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
       61 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
      121 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccggaa ttagatctct
      181 cgagatcgat gcggccgcgc caccatggaa caaaaactca tctcagaaga ggatctgatg
      241 gctgaaaaca aacaccctga caaaccactt aaggtgttgg aacagctggg caaagaagtc
      301 cttacggagt acctagaaaa attagtacaa agcaatgtac tgaaattaaa ggaggaagat
      361 aaacaaaaat ttaacaatgc tgaacgcagt gacaagcgtt gggtttttgt agatgccatg
      421 aaaaagaaac acagcaaagt aggtgaaatg cttctccaga cattcttcag tgtggaccca
      481 ggcagccacc atggtgaagc taatctggaa atggaggaac cagaagaatc attgaacact
      541 ctcaagcttt gttcccctga agagttcaca aggctttgca gagaaaagac acaagaaatt
      601 tacccaataa aggaggccaa tggccgtaca cgaaaggctc ttatcatatg caatacagag
      661 ttcaaacatc tctcactgag gtatggggct aaatttgaca tcattggtat gaaaggcctt
      721 cttgaagact taggctacga tgtggtggtg aaagaggagc ttacagcaga gggcatggag
      781 tcagagatga aagactttgc tgcactctca gaacaccaga catcagacag cacattcctg
      841 gtgctaatgt ctcatggcac actgcatggc atttgtggaa caatgcacag tgaaaaaact
      901 ccagatgtgc tacagtatga taccatctat cagatattca acaattgcca ctgtccaggt
      961 ctacgagaca aacccaaagt catcattgtg caggcctgca gaggtgggaa ctctggagaa
     1021 atgtggatca gagagtcttc aaaaccccag ttgtgcagag gtgtagatct acctaggaat
     1081 atggaagctg atgctgtcaa gctgagccac gtggagaagg acttcattgc cttctactct
     1141 acaaccccac atcacttgtc ctaccgagac aaaacaggag gctcttactt catcactaga
     1201 ctcatttcct gcttccggaa acatgcttgc tcttgtcatc tctttgatat attcctgaag
     1261 gtgcaacaat catttgaaaa ggcaagtatt cattcccaga tgcccaccat tgatcgggca
     1321 accttgacaa gatatttcta cctctttcct ggcaactgag tcgacggtat cgataagctt
     1381 gatatcgaat tccgcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     1441 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     1501 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     1561 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     1621 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     1681 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     1741 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     1801 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     1861 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     1921 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca
     1981 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2041 acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga
     2101 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2161 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2221 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2281 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2341 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2401 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2461 cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2521 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2581 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2641 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa tgaattaatt
     2701 aagaattcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     2761 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     2821 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     2881 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     2941 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3001 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     3061 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     3121 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct
     3181 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     3241 actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt
     3301 ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca
     3361 catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat
     3421 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3481 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3541 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3601 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3661 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4381 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4441 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4501 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5401 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5521 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5581 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     5641 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5701 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5761 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     5821 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     5881 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     5941 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6001 gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     6061 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6121 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6181 gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga
     6241 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
     6301 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6361 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6421 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     6481 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     6541 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     6601 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     6661 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     6721 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta ttcccaataa
     6781 agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg gtctcctcag
     6841 attgattgac tgcccacctc gggggtcttt catttggagg ttccaccgag atttggagac
     6901 ccctgcccag ggaccaccga cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt
     6961 gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta
     7021 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
     7081 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
     7141 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     7201 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     7261 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
     7321 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     7381 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     7441 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     7501 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     7561 gcatggacac ccagaccagg tcccctacat cgtgac
//
