LOCUS       sequence-433870-        7596 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 7596)
  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 7596)
  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-11_humanized-mutant
FEATURES             Location/Qualifiers
     source          1..7596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    578..919
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             986..1402
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1332..1354
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1370..1386
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1435..1444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1444..1473
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1721..1730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2627..3193
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2643..3195
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2787..2804)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3014..3033
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3207..3209,3210..3212,3213..3926)
                     /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(3252..3273)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3513..3532)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3860..3881
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3988..4502
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4664..4680
                     /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(4688..4694,4695..4712,4713..4718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4733..4754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4871..4888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5042..5630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5122..5141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5801..6592,6593..6661))
                     /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     6424..6443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6662..6766)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6834..6852
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6890..6912)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7012..7031
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7225..7247
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7375..7394
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7594..7596,1..514)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
       61 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
      121 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
      181 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
      241 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
      301 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
      361 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgta ttcccaataa
      421 agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg gtctcctcag
      481 attgattgac tgcccacctc gggggtcttt catttggagg ttccaccgag atttggagac
      541 ccctgcccag ggaccaccga cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt
      601 gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta
      661 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
      721 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
      781 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
      841 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
      901 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
      961 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     1021 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     1081 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     1141 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     1201 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     1261 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     1321 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     1381 cctcactcct tctctaggcg ccggaattag atctctcgag atcgatgcgg ccgcgccacc
     1441 atggaacaaa aactcatctc agaagaggat ctgatggctg aaaacaaaca ccctgacaaa
     1501 ccacttaagg tgttggaaca gctgggcaaa gaagtcctta cggagtacct agaaaaatta
     1561 gtacaaagca atgtactgaa attaaaggag gaagataaac aaaaatttaa caatgctgaa
     1621 cgcagtgaca agcgttgggt ttttgtagat gccatgaaaa agaaacacag caaagtaggt
     1681 gaaatgcttc tccagacatt cttcagtgtg gacccaggca gccaccatgg tgaagctaat
     1741 ctggaaatgg aggaaccaga agaatcattg aacactctca agctttgttc ccctgaagag
     1801 ttcacaaggc tttgcagaga aaagacacaa gaaatttacc caataaagga ggccaatggc
     1861 cgtacacgaa aggctcttat catatgcaat acagagttca aacatctctc actgaggtat
     1921 ggggctaaat ttgacatcat tggtatgaaa ggccttcttg aagacttagg ctacgatgtg
     1981 gtggtgaaag aggagcttac agcagagggc atggagtcag agatgaaaga ctttgctgca
     2041 ctctcagaac accagacatc agacagcaca ttcctggtgc taatgtctca tggcacactg
     2101 catggcattt gtggaacaat gcacagtgaa aaaactccag atgtgctaca gtatgatacc
     2161 atctatcaga tattcaacaa ttgccactgt ccaggtctac gagacaaacc caaagtcatc
     2221 attgtgcagg cctgcagagg tgggaactct ggagaaatgt ggatcagaga gtcttcaaaa
     2281 ccccagttgt gcagaggtgt agatctacct aggaatatgg aagctgatgc tgtctataaa
     2341 acccacgtgg agaaggactt cattgccttc tactctacaa ccccacatca cttgtcctac
     2401 cgagacaaaa caggaggctc ttacttcatc actagactca tttcctgctt ccggaaacat
     2461 gcttgctctt gtcatctctt tgatatattc ctgaaggtgc aacaatcatt tgaaactcca
     2521 agggccaaag ctcagatgcc caccattgat cgggcaacct tgacaagata tttctacctc
     2581 tttcctggca actgagtcga cggtatcgat aagcttgata tcgaattccg cccccccccc
     2641 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2701 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2761 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2821 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2881 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2941 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     3001 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     3061 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     3121 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3181 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3241 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtctg
     3301 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3361 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3421 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3481 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3541 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3601 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3661 atatcatggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
     3721 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3781 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3841 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3901 tcggcatgga cgagctgtac aagtaatgaa ttaattaaga attcgataaa ataaaagatt
     3961 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     4021 gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca
     4081 gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag
     4141 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca
     4201 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     4261 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     4321 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg
     4381 cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     4441 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     4501 atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa
     4561 gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag
     4621 ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat ccgctcacaa
     4681 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4741 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4801 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4861 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4921 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4981 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5041 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5101 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5161 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5221 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5281 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5341 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5401 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5461 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5521 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5581 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5641 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5701 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5761 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5821 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5881 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5941 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6001 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6061 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6121 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6181 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6241 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6301 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6361 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6421 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6481 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6541 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6601 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6661 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6721 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6781 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6841 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6901 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6961 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     7021 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     7081 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     7141 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     7201 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa
     7261 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     7321 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     7381 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     7441 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     7501 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     7561 aaagatttta tttagtctcc agaaaaaggg gggaat
//
