LOCUS       sequence-329618-        7380 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 7380)
  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 7380)
  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-TRE-DsRed-IRES-d24EGFP
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    98..116
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    140..158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    182..200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    224..242
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    266..284
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        317..355
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     352..376
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             549..1226
                     /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(679..697)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(729..749)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     947..966
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1141..1164
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    join(1269..1843,1844..1846,1847..1855)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1436..1453)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1663..1682
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1856..1858,1859..1861,1862..2575)
                     /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(1856..1858,1859..1861,1862..2572)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1901..1922)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2162..2181)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2509..2530
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2600..3016)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2616..2632)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2648..2670)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3083..3424
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3488..4004
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4204..4223)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4351..4373)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4567..4586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4686..4708
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4746..4764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4832..4936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4937..5005,5006..5797)
                     /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(5155..5174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5968..6556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6457..6476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6710..6727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6844..6865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6880..6885,6886..6903,6904..6910)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6918..6934
                     /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 ggttctctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac
       61 taaataaaat cttttatttt atcgtcgagt ttaccactcc ctatcagtga tagagaaaag
      121 tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
      181 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
      241 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagc
      301 tcggtacccg ggtcgaggta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
      361 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      421 ccgggaccga tccagcctcc gcggccccga attcgagctc ggtacccggg gatcctctag
      481 tcagctgacg cgtgctagcg cggccgcacc ggctagcgct accggactca gatccaccgg
      541 tcgccaccat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc aaggtgcgca
      601 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      661 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
      721 gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag caccccgccg
      781 acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      841 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggctcct
      901 tcatctacaa ggtgaagttc atcggcgtga acttcccctc cgacggcccc gtaatgcaga
      961 agaagactat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     1021 agggcgagat ccacaaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     1081 agtccatcta catggccaag aagcccgtgc agctgcccgg ctactactac gtggactcca
     1141 agctggacat cacctcccac aacgaggact acaccatcgt ggagcagtac gagcgcgccg
     1201 agggccgcca ccacctgttc ctgtagcggc caattctgca gtcgacggta ccgcgggccc
     1261 gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     1321 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     1381 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     1441 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     1501 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     1561 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     1621 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     1681 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     1741 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     1801 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
     1861 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1921 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1981 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2041 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2101 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2161 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2221 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2281 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2341 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2401 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2461 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2521 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     2581 cctcgagaga tctaattccg gcgcctagag aaggagtgag ggctggataa agggaggatc
     2641 gaggcggggt cgaacgagga ggttcaaggg ggagagacgg ggcggatgga ggaagaggag
     2701 gcggaggctt agggtgtaca aagggcttga cccagggagg ggggtcaaaa gccaaggctt
     2761 cccaggtcac gatgtagggg acctggtctg ggtgtccatg cgggccaggt gaaaagacct
     2821 tgatcttaac ctgggtgatg aggtctcggt taaaggtgcc gtctcgcggc catccgacgt
     2881 taaaggttgg ccattctgca gagcagaagg taacccaacg tctcttcttg acatctaccg
     2941 actggttgtg agcgatccgc tcgacatctt tccagtgacc taaggtcaaa cttaagggag
     3001 tggtaacagt ctggccctaa ttttcagaca aatacagaaa cacagtcaga cagagacaac
     3061 acagaacgat gctgcagcgc tgcagcagac aagacgcgcg gcttcggttc caaaccgaaa
     3121 gcaaaaattc agacggaggc gggaactgtt ttaggttctc gtctcctacc agaaccacat
     3181 atcctgacgg ggtcggattc cacatcgact cccttcctca ggtcgggcca caaaaacggc
     3241 ccccaaagtc cctgggacgt ctcccagggt tgcggccggg tgttcagaac tcgtcagttc
     3301 caccacgggt ccgccagata cagagctagt tagctaacta gtacagacgc aggcgcaaac
     3361 attagatgcc ggcacaaaca cgcacaaaga cagagacaga cacgaaacga ccgctggcca
     3421 gcttacctcc cggcgggggg gtcggtggtc cctgggcagg ggtctccaaa tctcggtgga
     3481 acctccaaat gaaagacccc cgaggtgggc agtcaatcaa tctgaggaga ccctcccaag
     3541 gatcagcgag tccacgattc ggatgcaaac agcaagaggc tttattggga atacgggtac
     3601 ccgggcgacg cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta
     3661 ttgagctcgg ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc
     3721 aaataaggca cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc
     3781 tagaaactgc tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg
     3841 gcaggaactg cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct
     3901 aaccttgatc tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg
     3961 ttacttaagc tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga
     4021 gactaaataa aatcttttat tttatctatg gctcgtactc tataggcttc agctggtgat
     4081 attgttgagt caaaactaga gcctggacca ctgatatcct gtctttaaca aattggacta
     4141 atcgcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     4201 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     4261 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     4321 tctccttgca tgcaccattc cttgcgccgt cgttttacaa cgtcgtgact gggaaaaccc
     4381 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4441 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4501 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     4561 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4621 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4681 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4741 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4801 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4921 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4981 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5041 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5101 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5161 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5221 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5281 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5341 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5401 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5461 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5521 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5581 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5641 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5701 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5761 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5941 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6001 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6061 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6541 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6601 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6661 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6721 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6781 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6841 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6901 tcgtatgttg tgtggaattg tgagcggata acaatttcac aattctcccc agctctttct
     6961 gcgctgtcca taatgaacta tttcaggagt attggagtat gtggattcaa aacagtggct
     7021 aattgagtca ggattactta atattcgact cctaccctca gaatgttgtt ctccaacttc
     7081 aagaaactgt tacccatgaa agacccccgc tgacgggtag tcaatcactc agaggagacc
     7141 ctcccaagga acagcgagac cacaagtcgg atgcaactgc aagagggttt attggataca
     7201 cgggtacccg ggcgacgcag tctatcggag gactggcgcg ccgagtgagg ggttgtgggc
     7261 tcttttattg agctcgggga gcagaagcgc gcgaacagaa gcgagaagcg aactgattgg
     7321 ttagttcaaa taaggcacag ggtcatttca ggtccttggg gcaccctgga aacatctgat
//
