LOCUS       sequence-361176-        7635 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 7635)
  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 7635)
  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-manatee-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7635
                     /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    1359..1925
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1375..1927
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1519..1536)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1746..1765
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1939..1941,1942..1944,1945..2658)
                     /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(1984..2005)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2245..2264)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2592..2613
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2720..3234
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3429..3445
                     /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(3453..3459,3460..3477,3478..3483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3498..3519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3636..3653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3807..4395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3887..3906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4566..5357,5358..5426))
                     /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     5189..5208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5427..5531)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5599..5617
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5655..5677)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5777..5796
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5990..6012
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6140..6159
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6359..6875
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6939..7280
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7347..7635,1..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 tggctgaaga caacaaagaa aacccattta aggttttcga
      241 atccttgggc aaagaaatat tgagtgacct tttggataat gtggaaaaaa aatacctgaa
      301 cttgtcgggc acagaaaagg aaaaatttta tgatgccaaa cctgcagaca agcttcgggt
      361 gttggaggcc tctgtccgaa agaagcgcga tgaggcaggc cgaatccttg tcgaaacgtt
      421 ctttagcaag gaaaaagagt ccaccagtgc agaagttctt ccagaaatca aggctggacc
      481 agatgaatca gcagaattca cagatcctct caagctctgt cctcgtgaag aattccagag
      541 actgtgtaaa gaaaaggctg gagagatcta tccaatcaaa gaaagaaaga accgcactcg
      601 tctggctctc attatatgca atatagagtt taacaatctc tctcggagga gtggggctga
      661 ggttgacatc gaagggatga agaggttgct tgagggcctg ggctacagcg tggatgtgaa
      721 aaaacaactc acagccacgg agatgcagtc agtgttgcag gaatttgctg cccgcccaga
      781 gcacaagtcc tcagacagca ctttcctggt gttcatgtct catggcttcc tggaaggaat
      841 atgtggaact atgcatagtg aagaacaacc agatatgtta tcctatgaca ccatcttcca
      901 gattttcaac aacaacaact gccagagtct aaaggacaaa ccgaagatta tcattgtcca
      961 ggcctgcaga ggtgaaaaga aaggaatatc gtggcacaag gactctctat cagtctcagc
     1021 agacagctct tcacagtcat ctgagaacct ggaagaggaa actattcaca acgcccacat
     1081 ggagaaagac ttcattgttt tctgggcctc aacaccacat aatttgtcct ggagagatga
     1141 aacacagggt tctcccttca tcacacgact catcacatgc ttacaaaaat attcttggtg
     1201 ctatcaccta gaagaagtat ttcagaaggt acaacaatcg tttgaaaaac catctattaa
     1261 aacccagatg cccactattg aacgggtcac tttgacaaga gatttctatc tctttcctgg
     1321 caattaagtc gacggtatcg ataagcttga tatcgaattc cgcccccccc cctaacgtta
     1381 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     1441 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     1501 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     1561 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     1621 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     1681 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     1741 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     1801 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     1861 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     1921 gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1981 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc
     2041 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2101 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2161 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2221 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2281 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2341 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2401 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
     2461 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2521 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2581 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2641 gacgagctgt acaagtaatg aattaattaa gaattcgata aaataaaaga ttttatttag
     2701 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     2761 aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg
     2821 ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct
     2881 gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc
     2941 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     3001 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     3061 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc
     3121 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     3181 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa
     3241 cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct
     3301 gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat
     3361 ggacagcgca gaaagagctg gggagaatwk trwrrwyrky rymsrmwsas mwkkssasam
     3421 twgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3481 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3541 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3601 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3781 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4441 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4501 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4561 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4621 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4681 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4741 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4801 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4861 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4921 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4981 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5041 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5101 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5161 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5221 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5281 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5341 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5401 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5461 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5521 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5581 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5641 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5701 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5761 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5821 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     5881 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     5941 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     6001 cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     6061 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     6121 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     6181 cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa tttgttaaag
     6241 acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     6301 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     6361 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     6421 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     6481 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6541 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     6601 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6661 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6721 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
     6781 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
     6841 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
     6901 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
     6961 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
     7021 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
     7081 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     7141 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     7201 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     7261 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     7321 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     7381 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     7441 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     7501 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     7561 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     7621 cccctccctg ggtca
//
