LOCUS       sequence-361030-        7605 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 7605)
  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 7605)
  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-bat-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     393..414
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    471..1023
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    473..1039
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(633..652)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     862..879
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2202..2231)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2231..2240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2273..2689)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2289..2305)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2321..2343)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2756..3097
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3161..3677
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3877..3896)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4024..4046)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4240..4259)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4359..4381
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4419..4437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4505..4609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4610..4678,4679..5470)
                     /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     complement(4828..4847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5641..6229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6130..6149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6383..6400
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6517..6538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6553..6558,6559..6576,6577..6583)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6591..6607
                     /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)."
     LTR             6769..7283
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(7345..7605,1..453,454..456,457..459))
                     /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(7390..7411)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
       61 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      121 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      181 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      241 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
      301 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
      361 ctcgccagac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
      421 caccaccccg gtgaacagct cctcgccctt gctcaccatg gttgtggcca tattatcatc
      481 gtgtttttca aaggaaaacc acgtccccgt ggttcggggg gcctagacgt ttttttaacc
      541 tcgactaaac acatgtaaag catgtgcacc gaggccccag atcagatccc atacaatggg
      601 gtaccttctg ggcatccttc agccccttgt tgaatacgct tgaggagagc catttgactc
      661 tttccacaac tatccaactc acaacgtggc actggggttg tgccgccttt gcaggtgtat
      721 cttatacacg tggcttttgg ccgcagaggc acctgtcgcc aggtgggggg ttccgctgcc
      781 tgcaaagggt cgctacagac gttgtttgtc ttcaagaagc ttccagagga actgcttcct
      841 tcacgacatt caacagacct tgcattcctt tggcgagagg ggaaagaccc ctaggaatgc
      901 tcgtcaagaa gacagggcca ggtttccggg ccctcacatt gccaaaagac ggcaatatgg
      961 tggaaaataa catatagaca aacgcacacc ggccttattc caagcggctt cggccagtaa
     1021 cgttaggggg gggggcggaa ttcgatatca agcttatcga taccgtcgac tcaattgcca
     1081 ggaaagaggt agaaatatct tgtcatggac tgtcgctcaa tagtgggcat ctgggcttta
     1141 acatctggtt tatcaaatga tcgttgtacc tttcgaaata cttcttctag gtgacagcac
     1201 caagaatagt tttgaaagca tgagatgagt tgtgcaatga agagagaacc ctttgtgata
     1261 tctctccagg acacattatg tggggtggag gagcagaaag caatgaagtc cttctccaca
     1321 tgtgtcttgt aaacggcatc ctcctccagg ttctcaagtg actgcgaatg gctgtctgcc
     1381 aaggctgctg gagagtcact gacccacaat tccccacgat ttgcacctct gcaggcctgg
     1441 acgatgatga cctttggttt gtcctttaga ctgaggcagt tgcggttgtt gaataacctg
     1501 aagatggtgt catatgggag cacatctggt ttttcatcac tatgcaaagt cccacagatt
     1561 ccatccagga tgccatgaga catgaatact aagaatgtgc tgtctgaaga cttgtgctct
     1621 gggcgggcag caaattcccg cagcactgac tccatttccg tggctgtgag tttctctttc
     1681 acatccacac tatagccaag gccctcaagc agctccttca tccctgtaac gtcaagctca
     1741 gaacccttcc tgagaggaag atgatcaaac tctgtattgc atatgatgag agccagacga
     1801 gtgcggtcct ttcgctcctt tattggatag atctctccag ccctttcttt acacagtttc
     1861 aggaactctt catgaggaca aagcttgagg gtatctgtag attctgctga ctcaacaggt
     1921 cccgccacta ttttcgaaag agcttctgta ctggtggaat ttttatccgt gttaaggaag
     1981 gtttggacaa ggacttgacc tgcctcgtgg cgtttctgtc gcacagagtc caccaagacc
     2041 caggccttgt ctgcaggttt ggcatcttga aattttttct tttcagtttc ctccagtttc
     2101 agaacatttt tttccaccaa gccatccaaa agtccagtta agatttcttt accgaaagtt
     2161 tccaaaacct taagtggttt tttgtcgtgg ttttcagcca tcagatcctc ttctgagatg
     2221 agtttttgtt ccatggtggc gcggccgcat cgatctcgag agatctaatt ccggcgccta
     2281 gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa
     2341 gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct
     2401 tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt
     2461 ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc
     2521 ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga
     2581 aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat
     2641 ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag
     2701 acaaatacag aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca
     2761 gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact
     2821 gttttaggtt ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg
     2881 actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag
     2941 ggttgcggcc gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct
     3001 agttagctaa ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa
     3061 agacagagac agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg
     3121 gtccctgggc aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg
     3181 ggcagtcaat caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca
     3241 aacagcaaga ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact
     3301 ggcgcgccga gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga
     3361 acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc
     3421 cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat
     3481 ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct
     3541 gtttggccca tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc
     3601 agttatgtat tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac
     3661 aggtggggtc tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct
     3721 atggctcgta ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga
     3781 ccactgatat cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg
     3841 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     3901 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     3961 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc
     4021 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4081 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4141 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     4201 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     4261 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4321 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4381 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4441 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4501 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4561 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4621 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4681 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4741 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4801 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4861 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4921 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4981 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5041 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5101 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5161 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5221 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5281 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5341 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5401 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5461 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5521 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5581 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5641 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5701 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5761 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5821 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5881 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5941 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6001 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6061 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6121 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6181 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6241 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6301 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6361 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6421 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6481 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6541 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6601 ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa ctatttcagg
     6661 agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac ttaatattcg
     6721 actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat gaaagacccc
     6781 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
     6841 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg cagtctatcg
     6901 gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     6961 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     7021 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggcggg
     7081 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     7141 atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc tgaacttctc
     7201 tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc tagcttgcca
     7261 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
     7321 tttatcgaat tcttaattaa ttcattactt gtacagctcg tccatgccga gagtgatccc
     7381 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     7441 cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     7501 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     7561 gatcttgaag ttcgccttga tgccgttctt ctgcttgtcg gccat
//
