LOCUS       sequence-361026-        7971 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 7971)
  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 7971)
  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-1/4b
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      161..170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             170..199
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1695..2261
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1711..2263
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1855..1872)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2082..2101
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2275..2277,2278..2280,2281..2994)
                     /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(2320..2341)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2581..2600)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2928..2949
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3056..3570
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3765..3781
                     /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(3789..3795,3796..3813,3814..3819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3834..3855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3972..3989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4143..4731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4223..4242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4902..5693,5694..5762))
                     /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     5525..5544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5763..5867)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5935..5953
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5991..6013)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6113..6132
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6326..6348
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6476..6495
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6695..7211
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7275..7616
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7683..7971,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct ctcgagatcg atgcggccgc gccaccatgg aacaaaaact
      181 catctcagaa gaggatctga tggccgacaa ggtcctgaag gagaagagaa agctgtttat
      241 ccgttccatg ggtgaaggta caataaatgg cttactggat gaattattac agacaagggt
      301 gctgaacaag gaagagatgg agaaagtaaa acgtgaaaat gctacagtta tggataagac
      361 ccgagctttg attgactccg ttattccgaa aggggcacag gcatgccaaa tttgcatcac
      421 atacatttgt gaagaagaca gttacctggc agggacgctg ggactctcag cagatcaaac
      481 atctggaaat taccttaata tgcaagactc tcaaggagta ctttcttcct ttccagaagg
      541 caaccacaga aaaaagccac ttaaggtgtt ggaatccctg ggcaaagatt tcctcactgg
      601 tgttttggat aacttggtgg aacaaaatgt actgaactgg aaggaagagg aaaaaaagaa
      661 atattacgat gctaaaactg aagacaaagt tcgggtcatg gcagactcta tgcaagagaa
      721 gcaacgtatg gcaggacaaa tgcttcttca aacctttttt aacatagacc aaatatcccc
      781 caataaaaaa gctcatccga atatggaggc tggaccacct gagtcaggag aatctacaga
      841 tgccctcaag ctttgtcctc atgaagaatt cctgagacta tgtaaagaaa gagctgaaga
      901 gatctatcca ataaaggaga gaaacaaccg cacacgcctg gctctcatca tatgcaatac
      961 agagtttgac catctgcctc cgaggaatgg agctgacttt gacatcacag ggatgaagga
     1021 gctacttgag ggtctggact atagtgtaga tgtagaagag aatctgacag ccagagatat
     1081 ggagtcagcg ctgagggcat ttgctaccag accagagcac aagtcctctg acagcacatt
     1141 cttggtactc atgtctcatg gcatcctgga gggaatctgc ggaactgtgc atgatgagaa
     1201 aaaaccagat gtgctgcttt atgacaccat cttccagata ttcaacaacc gcaactgcct
     1261 cagtctgaag gacaaaccca aggtcatcat tgtccaggcc tgcagaggtg caaaccgtgg
     1321 ggaactgtgg gtcagagact ctccagcatc cttggaagtg gcctcttcac agtcatctga
     1381 gaacctagag gaagatgctg tttacaagac ccacgtggag aaggacttca ttgctttctg
     1441 ctcttcaacg ccacacaacg tgtcctggag agacagcaca atgggctcta tcttcatcac
     1501 acaactcatc acatgcttcc agaaatattc ttggtgctgc cacctagagg aagtatttcg
     1561 gaaggtacag caatcatttg aaactccaag ggccaaagct caaatgccca ccatagaacg
     1621 actgtccatg acaagatatt tctacctctt tcctggcaat tgagtcgacg gtatcgataa
     1681 gcttgatatc gaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     1741 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     1801 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     1861 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     1921 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     1981 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2041 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     2101 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     2161 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2221 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     2281 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2341 gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag
     2401 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2461 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2521 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2581 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2641 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2701 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2761 aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2821 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2881 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2941 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt
     3001 aattaagaat tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     3061 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     3121 aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat
     3181 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3241 atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag
     3301 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     3361 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     3421 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc
     3481 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     3541 attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa
     3601 cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa
     3661 tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggka
     3721 gwwywktrwr rwyrkyrymg rmasagctgg ggagaattgt gaaattgtta tccgctcaca
     3781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3901 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5881 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5941 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6001 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6061 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     6121 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     6181 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6241 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6301 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
     6361 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     6421 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6481 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     6541 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     6601 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     6661 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     6721 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     6781 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     6841 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     6901 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     6961 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     7021 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     7081 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
     7141 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
     7201 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
     7261 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     7321 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     7381 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     7441 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     7501 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     7561 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     7621 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     7681 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     7741 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     7801 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     7861 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     7921 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc a
//
