LOCUS       sequence-422644-        7080 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 7080)
  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 7080)
  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-human-pro-IL-18_V47N_I48N
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             7..523
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    587..928
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             995..1411
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1341..1363
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1379..1395
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1444..1453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1453..1482
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2062..2085
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2120..2686
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2136..2688
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2280..2297)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2507..2526
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2700..2702,2703..2705,2706..3419)
                     /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(2745..2766)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3006..3025)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3353..3374
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3481..3995
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4157..4173
                     /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(4181..4187,4188..4205,4206..4211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4226..4247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4364..4381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4535..5123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4615..4634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5294..6085,6086..6154))
                     /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     5917..5936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6155..6259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6327..6345
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6383..6405)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6505..6524
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6718..6740
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6868..6887
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
       61 caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag
      121 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      181 ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc
      241 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
      301 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
      361 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtat
      421 tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg
      481 tctcctcaga ttgattgact gcccacctcg ggggtctttc atttggaggt tccaccgaga
      541 tttggagacc cctgcccagg gaccaccgac ccccccgccg ggaggtaagc tggccagcgg
      601 tcgtttcgtg tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct
      661 gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      721 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      781 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      841 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      901 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      961 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
     1021 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1081 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1141 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1201 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1261 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1321 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1381 ttatccagcc ctcactcctt ctctaggcgc cggaattaga tctctcgaga tcgatgcggc
     1441 cgcgccacca tggaacaaaa actcatctca gaagaggatc tgatggctgc tgaaccagta
     1501 gaagacaatt gcatcaactt tgtggcaatg aaatttattg acaatacgct ttactttata
     1561 gctgaagatg atgaaaacct ggaatcagat tactttggca agcttgaatc taaattatca
     1621 aataacagaa atttgaatga ccaagttctc ttcattgacc aaggaaatcg gcctctattt
     1681 gaagatatga ctgattctga ctgtagagat aatgcacccc ggaccatatt tattataagt
     1741 atgtataaag atagccagcc tagaggtatg gctgtaacta tctctgtgaa gtgtgagaaa
     1801 atttcaactc tctcctgtga gaacaaaatt atttccttta aggaaatgaa tcctcctgat
     1861 aacatcaagg atacaaaaag tgacatcata ttctttcaga gaagtgtccc aggacatgat
     1921 aataagatgc aatttgaatc ttcatcatac gaaggatact ttctagcttg tgaaaaagag
     1981 agagaccttt ttaaactcat tttgaaaaaa gaggatgaat tgggggatag atctataatg
     2041 ttcactgttc aaaacgaaga cgactacaag gacgacgatg acaagtaggt cgacggtatc
     2101 gataagcttg atatcgaatt ccgccccccc ccctaacgtt actggccgaa gccgcttgga
     2161 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2221 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2281 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2341 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2401 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2461 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2521 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2581 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     2641 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     2701 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2761 gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg
     2821 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2881 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2941 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3001 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3061 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3121 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3181 gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3241 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3301 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3361 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     3421 gaattaatta agaattcgat aaaataaaag attttattta gtctccagaa aaagggggga
     3481 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3541 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
     3601 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3661 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
     3721 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     3781 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     3841 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat
     3901 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     3961 gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag
     4021 aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt
     4081 ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct
     4141 ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5341 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5401 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5461 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5521 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5581 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5641 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5701 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5761 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5821 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5881 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5941 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6001 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6061 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6121 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6181 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6241 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6301 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     6361 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     6421 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6481 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6541 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     6601 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6661 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     6721 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6781 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6841 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     6901 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt
     6961 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct
     7021 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa
//
