LOCUS       sequence-361168-        7962 bp    DNA     circular SYN 15-APR-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 Apr 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-tiger-Caspase-1/4b
FEATURES             Location/Qualifiers
     source          1..7962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..122
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     138..154
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      203..212
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             212..241
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1761..2327
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1777..2329
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1921..1938)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2148..2167
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2341..2343,2344..2346,2347..3060)
                     /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(2386..2407)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2647..2666)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2994..3015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3122..3636
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3798..3814
                     /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(3822..3828,3829..3846,3847..3852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3867..3888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4005..4022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4176..4764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4256..4275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4935..5726,5727..5795))
                     /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     5558..5577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5796..5900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5968..5986
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6024..6046)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6146..6165
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6359..6381
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6509..6528
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6728..7244
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7308..7649
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7716..7962,1..170)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
       61 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
      121 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat
      181 ctctcgagat cgatgcggcc gcgccaccat ggaacaaaaa ctcatctcag aagaggatct
      241 gatggccgac aaggatctga agggcaagag gaagcagttc atcaactcag tcagcatggg
      301 gacggtcaac ggcttgctgg atgaactctt tgagaaaaat gtgctgaacc aggaggagat
      361 ggagagagtg aaatgtgaaa acgctacagt tatggataag gcccgagcgc tgatcgacag
      421 cgtcctgcga aaagggccac gggcgtgcca gatcgttatc tgtcacatct gtgaggaaga
      481 cacccacctt gcagagacca tggggctctc ctcaagccca caatctggaa attctcagaa
      541 caccacggac tctgaagtag cgtttcctcc tcttccagag gacaaacacg acaaaaaccc
      601 atttaagatt ttggaatctg tgggcaaaga gctccttact ggcgttttgc aagacttggt
      661 ggagaaagat gtcctgaaat tggaggaagc agaaaagaaa aaatttcatg atgccaaacc
      721 tggagacaag ccccgggtct ttctgaactc agtgcgtcag aaacccaagg aggcaggtca
      781 agccattgtc caaactttcc ttaatatgga caagcattct accagtttag aagctagcgt
      841 gaataacatg cctgggccgg ctgagccaga agaatctgta gatgctctca agctttgtcc
      901 tcgtgaaaac ttcgtgaaac tgtgtaaaca gagggctgaa gagatctacc caataaagga
      961 gagaaaggat cgtactcgtc tggctctcat catatgcaat acgacgttcg atcatctttc
     1021 tctcaggaag ggggctgacc ttgacgttgc agggatgagg aggctgctta cagaccttgg
     1081 ctacagtgtg cacgtaaagg aggaactcac tgctaaggac atggaatcag agctgagggc
     1141 atttgctgcc cgtccagagc acaagtcctc ggacagcaca ttcctggtgt tcatgtctca
     1201 tggcatcctg agtggaatct gtgggacgaa gtacagtgct gaaggagacc cagatgtatt
     1261 ggcttatgac accatcttcc agattttcaa caaccgcaac tgccttagtc taaaggacaa
     1321 gcccaaggtc atcatcgtcc aggcctgcag aggtgaaaat ttgggggaac tgttggtcag
     1381 tgactctcca gcggccccaa tggacagcac tacacagatg ggtagcagcc tttcacaggt
     1441 gggtgacaac ctagaggacg acgccgttta cagggtccac gtggagaagg acttcatcgc
     1501 tttctgctcc tcaaccccac atcatgtgtc ttggagagac gtgaacaagg gatctctctt
     1561 cattacacaa ctcatcacgt gcttccaaaa gtattcgtgg tgctctcatc tggaggaagt
     1621 atttcggaag gtacaacagt catttgaaaa accaaatgtt agagcccaga tgcccaccat
     1681 tgaacgacta tccatgacaa gatacttcta tctcttccct ggcaattgag tcgacggtat
     1741 cgataagctt gatatcgaat tccgcccccc cccctaacgt tactggccga agccgcttgg
     1801 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     1861 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     1921 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     1981 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2041 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2101 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2161 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2221 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     2281 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     2341 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2401 ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac
     2461 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2521 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2581 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2641 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2701 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2761 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2821 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2881 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2941 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3001 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3061 tgaattaatt aagaattcga taaaataaaa gattttattt agtctccaga aaaagggggg
     3121 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     3181 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
     3241 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     3301 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
     3361 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     3421 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     3481 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa
     3541 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     3601 tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga
     3661 gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt
     3721 tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc
     3781 tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3841 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4861 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4921 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4981 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5041 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5101 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5161 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5221 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5281 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5341 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5401 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5461 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5521 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5581 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5641 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5701 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5761 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5821 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5881 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5941 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6001 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     6061 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6121 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     6181 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     6241 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6301 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     6361 cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca
     6421 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     6481 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     6541 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat
     6601 ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc
     6661 tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa
     6721 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     6781 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     6841 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     6901 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     6961 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     7021 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     7081 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta
     7141 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg
     7201 gtctcctcag attgattgac tgcccacctc gggggtcttt catttggagg ttccaccgag
     7261 atttggagac ccctgcccag ggaccaccga cccccccgcc gggaggtaag ctggccagcg
     7321 gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc
     7381 tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
     7441 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
     7501 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
     7561 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
     7621 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
     7681 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
     7741 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     7801 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     7861 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     7921 cacctggccc gcatggacac ccagaccagg tcccctacat cg
//
