LOCUS       sequence-422629-        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
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             5..521
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    585..926
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             993..1409
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1339..1361
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1377..1393
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1442..1451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1451..1480
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2060..2083
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2118..2684
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2134..2686
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2278..2295)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2505..2524
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2698..2700,2701..2703,2704..3417)
                     /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(2743..2764)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3004..3023)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3351..3372
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3479..3993
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4155..4171
                     /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(4179..4185,4186..4203,4204..4209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4224..4245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4362..4379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4533..5121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4613..4632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5292..6083,6084..6152))
                     /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     5915..5934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6153..6257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6325..6343
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6381..6403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6503..6522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6716..6738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6866..6885
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
       61 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
      121 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
      181 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
      241 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
      301 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
      361 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
      421 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
      481 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
      541 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
      601 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
      661 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
      721 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
      781 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
      841 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
      901 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
      961 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     1021 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     1081 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     1141 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1201 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1261 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     1321 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     1381 atccagccct cactccttct ctaggcgccg gaattagatc tctcgagatc gatgcggccg
     1441 cgccaccatg gaacaaaaac tcatctcaga agaggatctg atggctgctg aaccagtaga
     1501 agacaattgc atcaactttg tggcaatgaa atttattgac aatacgcttt actttatagc
     1561 tgaagatgat gaaaacctgg aatcagatta ctttggcaag cttgaatcta aattatcagt
     1621 cataagaaat ttgaatgacc aagttctctt cattgaccaa ggaaatcggc ctctatttga
     1681 agatatgact gattctgact gtagagataa tgcaccccgg accatattta ttataagtat
     1741 gtataaagat agccagccta gaggtatggc tgtaactatc tctgtgaagt gtgagaaaat
     1801 ttcaactctc tcctgtgaga acaaaattat ttcctttaag gaaatgaatc ctcctgataa
     1861 catcaaggat acaaaaagtg acatcatatt ctttcagaga agtgtcccag gacatgataa
     1921 taagatgcaa tttgaatctt catcatacga aggatacttt ctagcttgtg aaaaagagag
     1981 agaccttttt aaactcattt tgaaaaaaga ggatgaattg ggggatagat ctataatgtt
     2041 cactgttcaa aacgaagacg actacaagga cgacgatgac aagtaggtcg acggtatcga
     2101 taagcttgat atcgaattcc gccccccccc ctaacgttac tggccgaagc cgcttggaat
     2161 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     2221 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     2281 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     2341 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     2401 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     2461 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2521 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2581 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2641 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     2701 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2761 gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc cacctacggc
     2821 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2881 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2941 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3001 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3061 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3121 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     3181 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3241 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3301 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3361 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatga
     3421 attaattaag aattcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3481 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3541 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     3601 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3661 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3721 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3781 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3841 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     3901 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3961 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     4021 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     4081 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     4141 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5221 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5281 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5341 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5401 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5461 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5521 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5581 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5641 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5701 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5761 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5821 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5881 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5941 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6001 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6061 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6121 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6181 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6241 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6301 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6361 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6421 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6481 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6541 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     6601 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6661 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6721 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     6781 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6841 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6901 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     6961 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     7021 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
//
