LOCUS       sequence-422635-        7608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 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, Dong Y, Mintseris J, Ma W, Gygi SP, Wu H, Kagan JC
  TITLE     Structural insights into cytokine cleavage by inflammatory 
            caspase-4.
  JOURNAL   Nature. 2023 Dec;624(7991):451-459. doi: 10.1038/s41586-023-06751-9.
            Epub 2023 Nov 22.
  PUBMED    37993712
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"; doi: 
            "10.1038/s41586-023-06751-9"; journalName: "Nature"; date: 
            "2023-12"; volume: "624"; issue: "7991"; pages: "451-459"
COMMENT     Sequence Label: pMSCV-Myc-Caspase-4_R269D
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    564..905
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             972..1388
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1340
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1356..1372
                     /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"
     CDS             1430..1459
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2625..3191
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2641..3193
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2785..2802)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3012..3031
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3205..3207,3208..3210,3211..3924)
                     /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(3250..3271)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3511..3530)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3858..3879
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3986..4500
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4662..4678
                     /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(4686..4692,4693..4710,4711..4716))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4731..4752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4869..4886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5040..5628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5120..5139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5799..6590,6591..6659))
                     /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     6422..6441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6660..6764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6832..6850
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6888..6910)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7010..7029
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7223..7245
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7373..7392
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7592..7608,1..500)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
       61 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
      121 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
      181 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
      241 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
      301 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
      361 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
      421 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
      481 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac
      541 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
      601 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
      661 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      721 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      781 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
      841 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
      901 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
      961 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1021 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1081 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1141 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1201 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1261 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1321 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1381 taggcgccgg aattagatct ctcgagatcg atgcggccgc gccaccatgg aacaaaaact
     1441 catctcagaa gaggatctga tggcagaagg caaccacaga aaaaagccac ttaaggtgtt
     1501 ggaatccctg ggcaaagatt tcctcactgg tgttttggat aacttggtgg aacaaaatgt
     1561 actgaactgg aaggaagagg aaaaaaagaa atattacgat gctaaaactg aagacaaagt
     1621 tcgggtcatg gcagactcta tgcaagagaa gcaacgtatg gcaggacaaa tgcttcttca
     1681 aacctttttt aacatagacc aaatatcccc caataaaaaa gctcatccga atatggaggc
     1741 tggaccacct gagtcaggag aatctacaga tgccctcaag ctttgtcctc atgaagaatt
     1801 cctgagacta tgtaaagaaa gagctgaaga gatctatcca ataaaggaga gaaacaaccg
     1861 cacacgcctg gctctcatca tatgcaatac agagtttgac catctgcctc cgaggaatgg
     1921 agctgacttt gacatcacag ggatgaagga gctacttgag ggtctggact atagtgtaga
     1981 tgtagaagag aatctgacag ccagggatat ggagtcagcg ctgagggcat ttgctaccag
     2041 accagagcac aagtcctctg acagcacatt cttggtactc atgtctcatg gcatcctgga
     2101 gggaatctgc ggaactgtgc atgatgagaa aaaaccagat gtgctgcttt atgacaccat
     2161 cttccagata ttcaacaacc gcaactgcct cagtctgaag gacaaaccca aggtcatcat
     2221 tgtccaggcc tgcagaggtg caaaccgtgg ggaactgtgg gtcgatgact ctccagcatc
     2281 cttggaagtg gcctcttcac agtcatctga gaacctggag gaagatgctg tttacaagac
     2341 ccacgtggag aaggacttca ttgctttctg ctcttcaacg ccacacaacg tgtcctggag
     2401 agacagcaca atgggctcta tcttcatcac acaactcatc acatgcttcc agaaatattc
     2461 ttggtgctgc cacctagagg aagtatttcg gaaggtacag caatcatttg aaactccaag
     2521 ggccaaagct caaatgccca ccatagaacg actgtccatg acaagatatt tctacctctt
     2581 tcctggcaat tgagtcgacg gtatcgataa gcttgatatc gaattccgcc ccccccccta
     2641 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     2701 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     2761 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     2821 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     2881 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     2941 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     3001 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     3061 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     3121 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     3181 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     3241 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     3301 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3361 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3421 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3481 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3541 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3601 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3661 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     3721 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3781 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3841 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3901 ggcatggacg agctgtacaa gtaatgaatt aattaagaat tcgataaaat aaaagatttt
     3961 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     4021 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     4081 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     4141 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     4201 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4261 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4321 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     4381 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4441 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     4501 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     4561 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     4621 cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt
     4681 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4741 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4801 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4861 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4921 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4981 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5041 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5101 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5161 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5221 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5281 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5341 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5401 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5461 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5521 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5581 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5641 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5701 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5761 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5821 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5881 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5941 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6001 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6061 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6121 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6181 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6241 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6301 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6361 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6421 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6481 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6541 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6601 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6661 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6721 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6781 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6841 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6901 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6961 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     7021 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7081 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     7141 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     7201 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg
     7261 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     7321 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     7381 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     7441 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     7501 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     7561 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacct
//
