LOCUS       sequence-422638-        7779 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 7779)
  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 7779)
  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-5
FEATURES             Location/Qualifiers
     source          1..7779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1436..1445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1445..1474
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1756..1767
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    2811..3377
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2827..3379
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2971..2988)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3198..3217
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3391..3393,3394..3396,3397..4110)
                     /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(3436..3457)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3697..3716)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4044..4065
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4172..4686
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4848..4864
                     /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(4872..4878,4879..4896,4897..4902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4917..4938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5055..5072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5226..5814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5306..5325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5985..6776,6777..6845))
                     /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     6608..6627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6846..6950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7018..7036
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7074..7096)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7196..7215
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7409..7431
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7559..7578
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7778..7779,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga gatcgatgcg gccgcgccac
     1441 catggaacaa aaactcatct cagaagagga tctgatggct gaagacagtg gcaaaaaaaa
     1501 aaggcgtaag aattttgaag ctatgttcaa aggtatcctt cagagtggat tggataactt
     1561 cgtgataaac cacatgctaa agaacaacgt ggctggacaa acatctatcc agaccctagt
     1621 acctaatacg gatcaaaagt caaccagtgt aaaaaaagac aaccacaaaa aaaaaacagt
     1681 taagatgttg gaatacctgg gcaaagatgt tcttcatggt gtttttaatt atttggcaaa
     1741 acacgatgtt ctgacattga aggaagagga aaagaaaaaa tattatgata ccaaaattga
     1801 agacaaggcc ctgatcttgg tagactcttt gcgaaagaat cgcgtggctc atcaaatgtt
     1861 tacccaaaca cttctcaata tggaccaaaa gatcaccagt gtaaaacctc ttctgcaaat
     1921 cgaggctgga ccacctgagt cagcagaatc tacaaatata ctcaaacttt gtcctcgtga
     1981 agaattcctg agactgtgta aaaaaaatca tgatgagatc tatccaataa aaaagagaga
     2041 ggaccgcaga cgcctggctc tcatcatatg caatacaaag tttgatcacc tgcctgcaag
     2101 gaatggggct cactatgaca tcgtggggat gaaaaggctg cttcaaggcc tgggctacac
     2161 tgtggttgac gaaaagaatc tcacagccag ggatatggag tcagtgctga gggcatttgc
     2221 tgccagacca gagcacaagt cctctgacag cacgttcttg gtactcatgt ctcatggcat
     2281 cctagaggga atctgcggaa ctgcgcataa aaagaaaaaa ccggatgtgc tgctttatga
     2341 caccatcttc cagatattca acaaccgcaa ctgcctcagt ctaaaggaca aacccaaggt
     2401 catcattgtc caggcctgca gaggtgaaaa acatggggaa ctctgggtca gagactctcc
     2461 agcatccttg gcactcatct cttcacagtc atctgagaac ctggaggcag attctgtttg
     2521 caagatccac gaggagaagg acttcattgc tttctgttct tcaacaccac ataacgtgtc
     2581 ctggagagac cgcacaaggg gctccatctt cattacggaa ctcatcacat gcttccagaa
     2641 atattcttgc tgctgccacc taatggaaat atttcggaag gtacagaaat catttgaagt
     2701 tccacaggct aaagcccaga tgcccaccat agaacgagca accttgacaa gagatttcta
     2761 cctctttcct ggcaattgag tcgacggtat cgataagctt gatatcgaat tccgcccccc
     2821 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2881 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2941 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     3001 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     3061 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     3121 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     3181 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     3241 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     3301 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     3361 gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga gctgttcacc
     3421 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3481 tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3541 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3601 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3661 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3721 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3781 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3841 gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac
     3901 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3961 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     4021 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4081 actctcggca tggacgagct gtacaagtaa tgaattaatt aagaattcga taaaataaaa
     4141 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     4201 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     4261 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     4321 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     4381 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     4441 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     4501 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     4561 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     4621 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     4681 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata
     4741 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa
     4801 atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc
     4861 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4921 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4981 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5041 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5101 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5161 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5221 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5281 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5341 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5401 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5461 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5521 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5581 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5641 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5701 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5761 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5821 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5881 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5941 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6001 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6061 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6121 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6181 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6241 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6301 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6361 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6421 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6481 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6541 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6601 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6661 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6721 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6781 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6841 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6901 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6961 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7021 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     7081 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     7141 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     7201 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     7261 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     7321 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     7381 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa
     7441 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     7501 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     7561 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     7621 tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg
     7681 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     7741 aataaaagat tttatttagt ctccagaaaa aggggggaa
//
