LOCUS       sequence-257023-        7928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Caspase-1 C285A with 2 copies of pLxIS motif from STING 
            fused to its N-ternimus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7928)
  AUTHORS   Tan Y, Kagan JC
  TITLE     Innate Immune Signaling Organelles Display Natural and Programmable 
            Signaling Flexibility.
  JOURNAL   Cell. 2019 Apr 4;177(2):384-398.e11. doi: 
            10.1016/j.cell.2019.01.039. Epub 2019 Mar 7.
  PUBMED    30853218
REFERENCE   2  (bases 1 to 7928)
  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"; doi: 
            "10.1016/j.cell.2019.01.039"; journalName: "Cell"; date: "2019-04-4-
            4"; volume: "177"; issue: "2"; pages: "384-398.e11"
COMMENT     Sequence Label: pMSCV-IRES-GFP-Caspase-1 (C285A) -NpLxIS
FEATURES             Location/Qualifiers
     source          1..7928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1421..1430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2962..3528
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2978..3530
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3122..3139)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3349..3368
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3542..3544,3545..3547,3548..4261)
                     /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(3587..3608)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3848..3867)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4195..4216
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4323..4837
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4999..5015
                     /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(5023..5029,5030..5047,5048..5053))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5068..5089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5206..5223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5377..5965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5457..5476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6136..6927,6928..6996))
                     /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     6759..6778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6997..7101)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7169..7187
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7225..7247)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7347..7366
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7560..7582
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7710..7729
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctgcc gccaccatgg ttaccatgaa
     1441 tgccccaatg acatcagttg cacctccccc tagtgtgctc agccaggagc ctcgactgct
     1501 gatttctggt atggaccagc ctcttccact taggaccgat ttgatcgtaa caatgaacgc
     1561 tcccatgacc agcgtcgcac caccacccag tgtcttgagc caggaacccc gactcctgat
     1621 tagtggtatg gatcaacccc tgccactgcg gacagacctc attgggggag gaggttccgg
     1681 agggggcggg tcaggagggg gcggtagtgc tgataagata ctgcgagcaa aaaggaagca
     1741 atttatcaat tcagtcagca ttgggacaat aaacggtctg ttggatgaac tcttggagaa
     1801 gcgggtcctt aatcaagagg agatggataa aattaagctc gctaatatta cagctatgga
     1861 caaggctagg gacttgtgtg accacgtctc caagaagggc cctcaggcca gccagatttt
     1921 cattacctat atttgcaatg aagattgtta cctggcaggt atacttgagc tgcaaagcgc
     1981 tccaagcgca gagacattcg tagccaccga agacagcaag ggaggtcacc ccagtagttc
     2041 tgagacaaag gaggagcaaa acaaagagga cggcaccttc ccaggactca ccgggaccct
     2101 gaagttttgt cctcttgaaa aagctcaaaa actctggaag gagaacccta gtgaaatcta
     2161 tcccattatg aatacaacca ccaggacccg gcttgccctc attatttgta acaccgagtt
     2221 ccaacacttg agtccacgcg tgggggcaca agttgacctc agagaaatga aattgttgct
     2281 tgaagacctc ggatataccg tgaaagtaaa ggagaacctt actgccttgg aaatggtgaa
     2341 agaggtcaaa gagtttgcag catgcccaga acacaagaca tccgactcca ctttcctggt
     2401 ttttatgagt cacgggatcc aggagggcat ctgtggtaca acttattcta atgaggtttc
     2461 agacatattg aaagtcgata ctatttttca aatgatgaat acactcaagt gtccttcttt
     2521 gaaggacaag ccaaaggtga taattattca agctgctcga ggcgagaaac agggtgtagt
     2581 actccttaaa gactccgttc gagatagtga ggaagatttt cttacagacg ctatatttga
     2641 ggacgacggt ataaagaagg ctcatattga gaaggatttc atagccttct gctcatctac
     2701 acctgataac gtctcatggc gacatcccgt acggggcagc ttgtttatcg agagccttat
     2761 caagcacatg aaggaatacg cttggagttg cgatctcgag gacatcttca gaaaggtacg
     2821 attctctttc gaacaacctg aattcagact gcagatgcct accgctgatc gggtaacatt
     2881 gactaagaga ttctaccttt ttccagggca ttaagcggcc gcgtttaaac gtcgacggta
     2941 tcgataagct tgatatcgaa ttccgccccc ccccctaacg ttactggccg aagccgcttg
     3001 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     3061 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     3121 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     3181 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     3241 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     3301 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     3361 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     3421 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     3481 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     3541 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     3601 cggcgacgta aacggccaca agttcagcgt gtctggcgag ggcgagggcg atgccaccta
     3661 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     3721 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     3781 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3841 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     3901 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     3961 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     4021 cggcatcaag gcgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     4081 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4141 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     4201 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     4261 atgaattaat taagaattcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     4321 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     4381 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     4441 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     4501 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     4561 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     4621 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     4681 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     4741 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     4801 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     4861 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     4921 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     4981 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5161 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7141 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     7201 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     7261 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     7321 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     7381 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     7441 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7501 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7561 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     7621 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     7681 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     7741 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     7801 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     7861 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     7921 aagggggg
//
