LOCUS       sequence-361178-        7623 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 7623)
  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 7623)
  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-wombat-Caspase-4
FEATURES             Location/Qualifiers
     source          1..7623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..87
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     103..119
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      168..177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             177..206
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1387..1953
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1403..1955
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1547..1564)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1774..1793
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1967..1969,1970..1972,1973..2686)
                     /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(2012..2033)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2273..2292)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2620..2641
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2748..3262
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3424..3440
                     /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(3448..3454,3455..3472,3473..3478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3493..3514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3631..3648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3802..4390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3882..3901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4561..5352,5353..5421))
                     /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     5184..5203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5422..5526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5594..5612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5650..5672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5772..5791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5985..6007
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6135..6154
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6354..6870
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6934..7275
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7342..7623,1..135)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
       61 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
      121 ccttctctag gcgccggaat tagatctctc gagatcgatg cggccgcgcc accatggaac
      181 aaaaactcat ctcagaagag gatctgatgg ctgataaaaa caccaagaat aatctattaa
      241 aggattttga attactggga aaggacatag tcaacaatgt tttggacaat ttgctggaaa
      301 gtgacatact gacattggag gaagacgaac aagagaagta ccgtgaagct caagctgtcg
      361 ccaaagctga aaagaaagct aaggtcctga taaattcttt cagtcggaaa ttccactccg
      421 cagggcaagt gttggccgaa agcctgttta acaaagatga acgggatgac agtgatgagg
      481 agtcattacc ttctgcttct gcagcaagcc aggccatgtc agctaagccc atgggtacac
      541 taaagctttg ttctccagaa gattttcaaa ggcagaggaa agagagggaa gcagagatat
      601 acccagtgaa ggagaaagaa ggccgtacac ggttggctct catcatctgt aacatagaat
      661 ttaaacatca ctctaagagg gccggagctg aagttgacat tgaaaggatg aaaatccttc
      721 ttgaagagct aggatacagt gtgaaaatcc taaagaacct cacatcttcg gaaatgaagt
      781 cagccctgag aaagtttgct gcccgccctg agcacatatc atctgatagc acttttgttg
      841 tgttcatgtc tcatggtctt ctggatagaa tttgtgggaa aaatcatcac aaagatgatc
      901 cagatttgct tgcttatgac accatattcc aaattctaaa cactaagaat tgcatcaatt
      961 tgaaagataa acccaaggtc atcataatcc aggcttgtcg tggactgaac catggattta
     1021 cgtgggcgag tgatgcccca gaaattgctg aagacagcca aagtcaactg caagtctttg
     1081 aggatgactc tgtccagaag gcccatttag aaaaggactt cattactttc tgctcctcaa
     1141 ctccacataa taagtcttgg agagacccta aaaatggctc catcttcatt atggagctca
     1201 tccattcttt ccaacagtat tcttggtgtt gccacataga acaaatcttc cgaaaggttc
     1261 agaatgcatt tgaaactccc aaagttaaga cccaaatgcc caccattgaa agactgtcca
     1321 tgaccagata tttctacctc tttccaggca attaagtcga cggtatcgat aagcttgata
     1381 tcgaattccg cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     1441 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     1501 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     1561 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     1621 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     1681 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     1741 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     1801 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     1861 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     1921 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     1981 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2041 ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2101 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2161 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2221 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2281 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2341 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2401 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggcgaa
     2461 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2521 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2581 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2641 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatgaa ttaattaaga
     2701 attcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     2761 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     2821 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     2881 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     2941 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     3001 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     3061 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     3121 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     3181 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     3241 cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga
     3301 gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata
     3361 ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg
     3421 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5581 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5641 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5701 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5761 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5821 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5881 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5941 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     6001 taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     6061 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6121 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     6181 gtgatgccgg ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg
     6241 atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa gcctatagag
     6301 tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     6361 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     6421 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
     6481 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6541 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
     6601 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     6661 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     6721 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc
     6781 tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg
     6841 attgactgcc cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct
     6901 gcccagggac caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct
     6961 gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag
     7021 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
     7081 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
     7141 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
     7201 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
     7261 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     7321 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     7381 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     7441 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     7501 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     7561 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     7621 ccc
//
