LOCUS       sequence-361152-        7608 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 7608)
  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 7608)
  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-redesigned-human-Caspase-4_ANR261ENP_D315H_M318K_T352
            K
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..95
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     111..127
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      176..185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             185..214
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1380..1946
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1396..1948
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1540..1557)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1767..1786
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1960..1962,1963..1965,1966..2679)
                     /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(2005..2026)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2266..2285)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2613..2634
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2741..3255
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3417..3433
                     /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(3441..3447,3448..3465,3466..3471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3486..3507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3624..3641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3795..4383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3875..3894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4554..5345,5346..5414))
                     /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     5177..5196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5415..5519)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5587..5605
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5643..5665)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5765..5784
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5978..6000
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6128..6147
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6347..6863
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6927..7268
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7335..7608,1..143)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
       61 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
      121 ccctcactcc ttctctaggc gccggaatta gatctctcga gatcgatgcg gccgcgccac
      181 catggaacaa aaactcatct cagaagagga tctgatggca gaaggcaacc acagaaaaaa
      241 gccacttaag gtgttggaat ccctgggcaa agatttcctc actggtgttt tggataactt
      301 ggtggaacaa aatgtactga actggaagga agaggaaaaa aagaaatatt acgatgctaa
      361 aactgaagac aaagttcggg tcatggcaga ctctatgcaa gagaagcaac gtatggcagg
      421 acaaatgctt cttcaaacct tttttaacat agaccaaata tcccccaata aaaaagctca
      481 tccgaatatg gaggctggac cacctgagtc aggagaatct acagatgccc tcaagctttg
      541 tcctcatgaa gaattcctga gactatgtaa agaaagagct gaagagatct atccaataaa
      601 ggagagaaac aaccgcacac gcctggctct catcatatgc aatacagagt ttgaccatct
      661 gcctccgagg aatggagctg actttgacat cacagggatg aaggagctac ttgagggtct
      721 ggactatagt gtagatgtag aagagaatct gacagccagg gatatggagt cagcgctgag
      781 ggcatttgct accagaccag agcacaagtc ctctgacagc acattcttgg tactcatgtc
      841 tcatggcatc ctggagggaa tctgcggaac tgtgcatgat gagaaaaaac cagatgtgct
      901 gctttatgac accatcttcc agatattcaa caaccgcaac tgcctcagtc tgaaggacaa
      961 acccaaggtc atcattgtcc aggcctgcag aggtgaaaac ccgggggaac tgtgggtcag
     1021 agactctcca gcatccttgg aagtggcctc ttcacagtca tctgagaacc tagaggaaga
     1081 tgctgtttac aagacccacg tggagaagga cttcattgct ttctgctctt caacgccaca
     1141 caacgtgtcc tggagacaca gcacaaaagg ctctatcttc atcacacaac tcatcacatg
     1201 cttccagaaa tattcttggt gctgccacct agaggaagta tttcggaagg tacagcaatc
     1261 atttgaaaaa ccaagggcca aagctcaaat gcccaccata gaacgactgt ccatgacaag
     1321 atatttctac ctctttcctg gcaattgagt cgacggtatc gataagcttg atatcgaatt
     1381 ccgccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     1441 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     1501 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     1561 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     1621 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     1681 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     1741 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     1801 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     1861 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     1921 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     1981 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2041 ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2101 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2161 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2221 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2281 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2341 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2401 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggc gaacttcaag
     2461 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2521 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2581 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2641 gccgggatca ctctcggcat ggacgagctg tacaagtaat gaattaatta agaattcgat
     2701 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     2761 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     2821 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     2881 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     2941 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     3001 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     3061 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     3121 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     3181 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     3241 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     3301 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     3361 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     3421 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3481 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3541 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3601 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3661 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3721 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3781 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3841 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3901 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3961 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4021 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4081 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4141 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4201 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4261 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4321 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4381 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4441 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4501 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4561 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4621 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4681 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4741 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4801 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4861 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4921 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4981 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5041 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5101 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5161 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5221 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5281 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5341 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5401 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5461 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5521 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5581 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5641 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5701 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5761 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5821 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5881 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5941 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     6001 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     6061 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6121 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6181 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     6241 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     6301 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     6361 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     6421 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     6481 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     6541 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6601 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     6661 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     6721 gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct
     6781 gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact
     6841 gcccacctcg ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg
     6901 gaccaccgac ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg
     6961 tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta
     7021 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     7081 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     7141 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     7201 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
     7261 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     7321 tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga
     7381 aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta
     7441 ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta
     7501 accgagacct catcacccag gttaagatca aggtcttttc acctggcccg catggacacc
     7561 cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacc
//
