LOCUS       sequence-361117-        7962 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 7962)
  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 7962)
  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-cheetah-Caspase-1/4b
FEATURES             Location/Qualifiers
     source          1..7962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..125
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     141..157
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      206..215
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             215..244
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1764..2330
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1780..2332
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1924..1941)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2151..2170
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2344..2346,2347..2349,2350..3063)
                     /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(2389..2410)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2650..2669)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2997..3018
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3125..3639
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3801..3817
                     /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(3825..3831,3832..3849,3850..3855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3870..3891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4008..4025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4179..4767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4259..4278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4938..5729,5730..5798))
                     /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     5561..5580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5799..5903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5971..5989
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6027..6049)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6149..6168
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6362..6384
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6512..6531
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6731..7247
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7311..7652
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7719..7962,1..173)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
       61 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
      121 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaatta
      181 gatctctcga gatcgatgcg gccgcgccac catggaacaa aaactcatct cagaagagga
      241 tctgatggcc gacaaggatc tgaagggcaa gaggaagcag ttcatcaact cagtcggcat
      301 ggggacggtc aacggcttgc tggatgaact ctttgagaaa aatgtgctga accaggagga
      361 gatggagaga gtaaaatgtg aaaacgctac cgttatggat aaggcccgag ctctgatcga
      421 cagcgtcctg cgaaaagggc cacgggcgtg ccagatcttt atctgtcaca tctgtgagga
      481 agacacccac cttgcggaga cgctggggct gtcctcaagc ccacaatctg gaaattctca
      541 gaacaccacg gactctgaag tagcgtttcc tcctcttcca gaggacaaac acgaccaaaa
      601 cccatttaag attttggaat ctgtgggcaa agtgctcctt actggcgttt tgcaagactt
      661 ggtggagaaa gatgtcctga aattggagga agcagaaaag aaaaaatttc atgatgccaa
      721 acctggagac aagccccggg tctttctgaa ctcagtgcgt cagaaaccca aggaggcagg
      781 tcaagccatt gtccaaactt tccttaatac ggacaagcat tctaccagtt tagaagctag
      841 cgtgaataac atgcctgggc cggctgagcc agaagaatct gtagatgctc tcaagctttg
      901 tcctcgtgaa aacttcgtga aactgtgtaa acagagggct gaagagatct acccaataaa
      961 ggagagaaag gatcgtactc gtctggctct catcatctgc aatacgacgt tcgatcatct
     1021 ttctctcagg aagggggctg acctcgatgt tgcagggatg aggaggctgc ttacagacct
     1081 tggctacagt gtgcacataa aagaggaact cactgctaag gacatggaat cagagctgag
     1141 ggcatttgct gcccgtccag agcacaagtc ctcggacagc acattcctgg tgttcatgtc
     1201 tcatggcatc ctgagtggaa tctgtgggac gaagtacagc gctgaaggag acccagatgt
     1261 attggcttat gacaccatct tccagatttt caacaaccgc aactgcctta gtctaaagga
     1321 caagcccaag gtcatcatcg tccaggcctg cagaggtgaa aatttggggg aactgttgat
     1381 cagtgactct ccagcggccc caatggacag cacttcacag atgggtagca gcctttcaca
     1441 ggtgggtgac aacctagagg acgacgccat ttacaaggtc cacgtggaga aggacttcat
     1501 cgctttctgc tcctcgaccc cacatcatgt gtcttggaga gacgtgaaca agggatctct
     1561 cttcattaca caactcatca cgtgcttcca aaagtattcg tggtgctctc atctggagga
     1621 agtatttcgg aaggtacaac agtcatttga aaaaccaaat gttagagccc agatgcccac
     1681 cattgaacga ctatccatga caagatactt ctatctcttc cctggcaatt gagtcgacgg
     1741 tatcgataag cttgatatcg aattccgccc ccccccctaa cgttactggc cgaagccgct
     1801 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     1861 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     1921 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     1981 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     2041 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     2101 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     2161 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     2221 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     2281 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     2341 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2401 gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc
     2461 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2521 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2581 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2641 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2701 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2761 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2821 aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2881 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2941 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3001 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3061 taatgaatta attaagaatt cgataaaata aaagatttta tttagtctcc agaaaaaggg
     3121 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     3181 ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg aacagagaga
     3241 cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     3301 caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga
     3361 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     3421 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     3481 aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc
     3541 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     3601 ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt
     3661 ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc aattagccac
     3721 tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag
     3781 agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3841 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3901 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3961 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4021 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4081 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4141 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4201 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4261 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4321 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4381 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4441 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4501 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4561 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4621 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4681 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4741 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4801 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4861 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4921 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4981 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5041 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5101 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5161 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5221 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5281 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5341 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5401 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5461 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5521 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5581 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5641 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5701 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5761 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5821 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5881 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5941 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6001 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6061 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6121 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6181 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6241 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6301 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6361 tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc
     6421 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6481 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6541 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc
     6601 aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc
     6661 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga
     6721 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     6781 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     6841 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     6901 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     6961 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     7021 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     7081 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     7141 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg
     7201 agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc
     7261 gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca
     7321 gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc
     7381 gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
     7441 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
     7501 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
     7561 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     7621 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
     7681 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     7741 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     7801 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     7861 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     7921 tttcacctgg cccgcatgga cacccagacc aggtccccta ca
//
