LOCUS       sequence-329613-        7741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of DsRed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7741)
  AUTHORS   Yen HC, Xu Q, Chou DM, Zhao Z, Elledge SJ
  TITLE     Global protein stability profiling in mammalian cells.
  JOURNAL   Science. 2008 Nov 7;322(5903):918-23. doi: 10.1126/science.1160489.
  PUBMED    18988847
REFERENCE   2  (bases 1 to 7741)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2008-11-7- 7"; volume: "322"; issue: 
            "5903"; pages: "918-23"
COMMENT     Sequence Label: MSCV-CMV-DsRed-IRES-d4EGFP
FEATURES             Location/Qualifiers
     source          1..7741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             688..1365
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(818..836)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(868..888)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1086..1105
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1280..1303
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    join(1408..1982,1983..1985,1986..1994)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1575..1592)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1802..1821
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1995..1997,1998..2000,2001..2711,2712..2717,
                     2718..2840)
                     /codon_start=1
                     /product="EGFP destabilized by a mutated version of 
                     residues 422-461 of mouse ornithine decarboxylase, giving 
                     an in vivo half-life of ~4 hours"
                     /label=d4EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGALPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(2040..2061)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2301..2320)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2648..2669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2864..3280)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2880..2896)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2912..2934)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3347..3688
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3752..4268
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4468..4487)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4615..4637)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4831..4850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4950..4972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5010..5028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5096..5200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5201..5269,5270..6061)
                     /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     complement(5419..5438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6232..6820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6721..6740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6974..6991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7108..7129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7144..7149,7150..7167,7168..7174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7182..7198
                     /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)."
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
      601 ggaacggtgc attggaacgg acctgcagga attgatccgc ggccgcaccg gctagcgcta
      661 ccggactcag atccaccggt cgccaccatg gcctcctccg aggacgtcat caaggagttc
      721 atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      781 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
      841 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
      901 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
      961 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1021 tccctgcagg acggctcctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc
     1081 gacggccccg taatgcagaa gaagactatg ggctgggagg cctccaccga gcgcctgtac
     1141 ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa ggacggcggc
     1201 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc
     1261 tactactacg tggactccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     1321 gagcagtacg agcgcgccga gggccgccac cacctgttcc tgtagcggcc aattctgcag
     1381 tcgacggtac cgcgggcccg ggatccgccc ctctccctcc ccccccccta acgttactgg
     1441 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     1501 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     1561 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     1621 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     1681 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     1741 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     1801 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     1861 tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa
     1921 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     1981 atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2041 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2101 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2161 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2221 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2281 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2341 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2401 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2461 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2521 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2581 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2641 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2701 agctgtacaa gaagcttagc catggcttcc cgccggaggt ggaggagcag gatgatggcg
     2761 cgctgcccat gtcttgtgcc caggagagcg ggatggaccg tcaccctgca gcctgtgctt
     2821 ctgctaggat caatgtgtag gcggcctcga gagatctaat tccggcgcct agagaaggag
     2881 tgagggctgg ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag
     2941 acggggcgga tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg
     3001 gaggggggtc aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc
     3061 catgcgggcc aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg
     3121 tgccgtctcg cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc
     3181 aacgtctctt cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt
     3241 gacctaaggt caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca
     3301 gaaacacagt cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg
     3361 cgcggcttcg gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt
     3421 tctcgtctcc taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc
     3481 ctcaggtcgg gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc
     3541 cgggtgttca gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta
     3601 actagtacag acgcaggcgc aaacattaga tgccggcaca aacacgcaca aagacagaga
     3661 cagacacgaa acgaccgctg gccagcttac ctcccggcgg gggggtcggt ggtccctggg
     3721 caggggtctc caaatctcgg tggaacctcc aaatgaaaga cccccgaggt gggcagtcaa
     3781 tcaatctgag gagaccctcc caaggatcag cgagtccacg attcggatgc aaacagcaag
     3841 aggctttatt gggaatacgg gtacccgggc gacgcagtct atcggaggac tggcgcgccg
     3901 agtgaggggt tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg
     3961 agaagcgaac tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca
     4021 ccctggaaac atctgatggt tctctagaaa ctgctgaggg cgggaccgca tctggggacc
     4081 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
     4141 atattctgct gtctctctgt tcctaacctt gatctgaact tctctattct cagttatgta
     4201 ttttccatgc cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt
     4261 ctttcattcc cccctttttc tggagactaa ataaaatctt ttattttatc tatggctcgt
     4321 actctatagg cttcagctgg tgatattgtt gagtcaaaac tagagcctgg accactgata
     4381 tcctgtcttt aacaaattgg actaatcgcc tctacgccgg acgcatcgtg gccggcatca
     4441 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     4501 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     4561 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg ccgtcgtttt
     4621 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4681 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4741 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4801 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4861 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4921 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4981 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5041 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5161 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5221 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5281 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5341 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5401 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5461 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5521 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5581 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5641 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5701 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5761 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5821 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5881 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5941 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6001 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6061 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6121 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6181 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6241 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6301 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6361 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6421 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6481 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6541 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6601 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6661 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6721 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6781 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6841 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6901 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6961 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7021 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7081 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7141 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7201 tcacaattct ccccagctct ttctgcgctg tccataatga actatttcag gagtattgga
     7261 gtatgtggat tcaaaacagt ggctaattga gtcaggatta cttaatattc gactcctacc
     7321 ctcagaatgt tgttctccaa cttcaagaaa ctgttaccca tgaaagaccc ccgctgacgg
     7381 gtagtcaatc actcagagga gaccctccca aggaacagcg agaccacaag tcggatgcaa
     7441 ctgcaagagg gtttattgga tacacgggta cccgggcgac gcagtctatc ggaggactgg
     7501 cgcgccgagt gaggggttgt gggctctttt attgagctcg gggagcagaa gcgcgcgaac
     7561 agaagcgaga agcgaactga ttggttagtt caaataaggc acagggtcat ttcaggtcct
     7621 tggggcaccc tggaaacatc tgatggttct ctagcttgcc aaacctacag gtggggtctt
     7681 tcattccccc ctttttctgg agactaaata aaatctttta ttttatcgat cctctagagt
     7741 c
//
