LOCUS       sequence-329616-        7616 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 7616)
  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 7616)
  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-d24EGFP
FEATURES             Location/Qualifiers
     source          1..7616
                     /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..2714)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1995..1997,1998..2000,2001..2711)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     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(2739..3155)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2755..2771)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2787..2809)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3222..3563
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3627..4143
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4343..4362)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4490..4512)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4706..4725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4825..4847
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4885..4903)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4971..5075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5076..5144,5145..5936)
                     /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(5294..5313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6107..6695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6596..6615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6849..6866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6983..7004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7019..7024,7025..7042,7043..7049)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7057..7073
                     /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 gtaaagcggc ctcgagagat ctaattccgg cgcctagaga aggagtgagg
     2761 gctggataaa gggaggatcg aggcggggtc gaacgaggag gttcaagggg gagagacggg
     2821 gcggatggag gaagaggagg cggaggctta gggtgtacaa agggcttgac ccagggaggg
     2881 gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga cctggtctgg gtgtccatgc
     2941 gggccaggtg aaaagacctt gatcttaacc tgggtgatga ggtctcggtt aaaggtgccg
     3001 tctcgcggcc atccgacgtt aaaggttggc cattctgcag agcagaaggt aacccaacgt
     3061 ctcttcttga catctaccga ctggttgtga gcgatccgct cgacatcttt ccagtgacct
     3121 aaggtcaaac ttaagggagt ggtaacagtc tggccctaat tttcagacaa atacagaaac
     3181 acagtcagac agagacaaca cagaacgatg ctgcagcgct gcagcagaca agacgcgcgg
     3241 cttcggttcc aaaccgaaag caaaaattca gacggaggcg ggaactgttt taggttctcg
     3301 tctcctacca gaaccacata tcctgacggg gtcggattcc acatcgactc ccttcctcag
     3361 gtcgggccac aaaaacggcc cccaaagtcc ctgggacgtc tcccagggtt gcggccgggt
     3421 gttcagaact cgtcagttcc accacgggtc cgccagatac agagctagtt agctaactag
     3481 tacagacgca ggcgcaaaca ttagatgccg gcacaaacac gcacaaagac agagacagac
     3541 acgaaacgac cgctggccag cttacctccc ggcggggggg tcggtggtcc ctgggcaggg
     3601 gtctccaaat ctcggtggaa cctccaaatg aaagaccccc gaggtgggca gtcaatcaat
     3661 ctgaggagac cctcccaagg atcagcgagt ccacgattcg gatgcaaaca gcaagaggct
     3721 ttattgggaa tacgggtacc cgggcgacgc agtctatcgg aggactggcg cgccgagtga
     3781 ggggttgtgg gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag
     3841 cgaactgatt ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg
     3901 gaaacatctg atggttctct agaaactgct gagggcggga ccgcatctgg ggaccatctg
     3961 ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt
     4021 ctgctgtctc tctgttccta accttgatct gaacttctct attctcagtt atgtattttc
     4081 catgccttgc aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc
     4141 attcccccct ttttctggag actaaataaa atcttttatt ttatctatgg ctcgtactct
     4201 ataggcttca gctggtgata ttgttgagtc aaaactagag cctggaccac tgatatcctg
     4261 tctttaacaa attggactaa tcgcctctac gccggacgca tcgtggccgg catcaccggc
     4321 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
     4381 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     4441 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcgccgtc gttttacaac
     4501 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4561 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4621 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4681 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4741 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4801 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4861 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4921 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4981 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5041 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5101 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5161 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5221 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5281 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5341 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5401 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5461 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5521 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5581 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5641 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5701 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5761 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5821 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5881 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5941 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6001 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6061 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6181 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6241 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6481 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6721 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6781 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6841 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6901 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6961 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     7021 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     7081 attctcccca gctctttctg cgctgtccat aatgaactat ttcaggagta ttggagtatg
     7141 tggattcaaa acagtggcta attgagtcag gattacttaa tattcgactc ctaccctcag
     7201 aatgttgttc tccaacttca agaaactgtt acccatgaaa gacccccgct gacgggtagt
     7261 caatcactca gaggagaccc tcccaaggaa cagcgagacc acaagtcgga tgcaactgca
     7321 agagggttta ttggatacac gggtacccgg gcgacgcagt ctatcggagg actggcgcgc
     7381 cgagtgaggg gttgtgggct cttttattga gctcggggag cagaagcgcg cgaacagaag
     7441 cgagaagcga actgattggt tagttcaaat aaggcacagg gtcatttcag gtccttgggg
     7501 caccctggaa acatctgatg gttctctagc ttgccaaacc tacaggtggg gtctttcatt
     7561 cccccctttt tctggagact aaataaaatc ttttatttta tcgatcctct agagtc
//
