LOCUS       sequence-366973-        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane-targeted mCherry expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  AUTHORS   Yang JM, Chi WY, Liang J, Takayanagi S, Iglesias PA, Huang CH
  TITLE     Deciphering cell signaling networks with massively multiplexed 
            biosensor barcoding.
  JOURNAL   Cell. 2021 Dec 9;184(25):6193-6206.e14. doi: 
            10.1016/j.cell.2021.11.005. Epub 2021 Nov 26.
  PUBMED    34838160
REFERENCE   2  (bases 1 to 7292)
  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: 
            "10.1016/j.cell.2021.11.005"; journalName: "Cell"; date: "2021-12-9-
            9"; volume: "184"; issue: "25"; pages: "6193-6206.e14"
COMMENT     Sequence Label: pVITRO1-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..7292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..276
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(108..128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        290..1596
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          617..1564
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1641..1762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1652..1671)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1706..1725
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1950..2538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2439..2458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2618..2921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3036..4341
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3378..4309
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      4348..4357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4354..5061
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4499..4517)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4767..4786
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5173..5617
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5650..5696
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5688..5706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5716..6741
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    join(6753..7292,1..33)
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 caaaaactga gtccttgggt tgcatagaaa gctgcctgca gggcctgaaa taacctctga
       61 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
      121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      241 aagcatgcat ctcaattagt cagcaaccat agtcccacta gtggagccga gagtaattca
      301 tacaaaagga gggatcgcct tcgcaagggg agagcccagg gaccgtccct aaattctcac
      361 agacccaaat ccctgtagcc gccccacgac agcgcgagga gcatgcgctc agggctgagc
      421 gcggggagag cagagcacac aagctcatag accctggtcg tgggggggag gaccggggag
      481 ctggcgcggg gcaaactggg aaagcggtgt cgtgtgctgg ctccgccctc ttcccgaggg
      541 tgggggagaa cggtatataa gtgcggcagt cgccttggac gttctttttc gcaacgggtt
      601 tgccgtcaga acgcaggtga ggggcgggtg tggcttccgc gggccgccga gctggaggtc
      661 ctgctccgag cgggccgggc cccgctgtcg tcggcgggga ttagctgcga gcattcccgc
      721 ttcgagttgc gggcggcgcg ggaggcagag tgcgaggcct agcggcaacc ccgtagcctc
      781 gcctcgtgtc cggcttgagg cctagcgtgg tgtccgcgcc gccgccgcgt gctactccgg
      841 ccgcactctg gtcttttttt tttttgttgt tgttgccctg ctgccttcga ttgccgttca
      901 gcaatagggg ctaacaaagg gagggtgcgg ggcttgctcg cccggagccc ggagaggtca
      961 tggttgggga ggaatggagg gacaggagtg gcggctgggg cccgcccgcc ttcggagcac
     1021 atgtccgacg ccacctggat ggggcgaggc ctggggtttt tcccgaagca accaggctgg
     1081 ggttagcgtg ccgaggccat gtggccccag cacccggcac gatctggctt ggcggcgccg
     1141 cgttgccctg cctccctaac tagggtgagg ccatcccgtc cggcaccagt tgcgtgcgtg
     1201 gaaagatggc cgctcccggg ccctgttgca aggagctcaa aatggaggac gcggcagccc
     1261 ggtggagcgg gcgggtgagt cacccacaca aaggaagagg gcctggtccc tcaccggctg
     1321 ctgcttcctg tgaccccgtg gtcctatcgg ccgcaatagt cacctcgggc ttttgagcac
     1381 ggctagtcgc ggcgggggga ggggatgtaa tggcgttgga gtttgttcac atttggtggg
     1441 tggagactag tcaggccagc ctggcgctgg aagtcatttt tggaatttgt ccccttgagt
     1501 tttgagcgga gctaattctc gggcttctta gcggttcaaa ggtatctttt aaaccctttt
     1561 ttaggtgttg tgaaaaccac cgctaattca aagcaaccgg tgatatcaaa gatcttgtac
     1621 agctagctgg ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1681 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1741 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1801 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggaaat
     1861 gttaattaac tagccatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1921 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1981 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2041 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2101 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2161 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2221 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2281 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2341 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2401 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2461 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2521 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2581 tggccttttg ctcacatgtt cttaattaac ctgcaggcgt tacataactt acggtaaatg
     2641 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2701 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2761 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2821 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2881 cttggcagta catctacgta ttagtcatcg ctattaccat gatgatgcgg ttttggcagt
     2941 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     3001 acgtcaatgg gagtttgttt tgactagtgg agccgagagt aattcataca aaaggaggga
     3061 tcgccttcgc aaggggagag cccagggacc gtccctaaat tctcacagac ccaaatccct
     3121 gtagccgccc cacgacagcg cgaggagcat gcgcccaggg ctgagcgcgg gtagatcaga
     3181 gcacacaagc tcacagtccc cggcggtggg gggaggggcg cgctgagcgg gggccaggga
     3241 gctggcgcgg ggcaaactgg gaaagtggtg tcgtgtgctg gctccgccct cttcccgagg
     3301 gtgggggaga acggtatata agtgcggtag tcgccttgga cgttcttttt cgcaacgggt
     3361 ttgccgtcag aacgcaggtg agtggcgggt gtggcttccg cgggccccgg agctggagcc
     3421 ctgctctgag cgggccgggc tgatatgcga gtgtcgtccg cagggtttag ctgtgagcat
     3481 tcccacttcg agtggcgggc ggtgcggggg tgagagtgcg aggcctagcg gcaaccccgt
     3541 agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc cgccgccgcg tgccactccg
     3601 gccgcactat gcgttttttg tccttgctgc cctcgattgc cttccagcag catgggctaa
     3661 caaagggagg gtgtggggct cactcttaag gagcccatga agcttacgtt ggataggaat
     3721 ggaagggcag gaggggcgac tggggcccgc ccgccttcgg agcacatgtc cgacgccacc
     3781 tggatggggc gaggcctgtg gctttccgaa gcaatcgggc gtgagtttag cctacctggg
     3841 ccatgtggcc ctagcactgg gcacggtctg gcctggcggt gccgcgttcc cttgcctccc
     3901 aacaagggtg aggccgtccc gcccggcacc agttgcttgc gcggaaagat ggccgctccc
     3961 ggggccctgt tgcaaggagc tcaaaatgga ggacgcggca gcccggtgga gcgggcgggt
     4021 gagtcaccca cacaaaggaa gagggccttg cccctcgccg gccgctgctt cctgtgaccc
     4081 cgtggtctat cggccgcata gtcacctcgg gcttctcttg agcaccgctc gtcgcggcgg
     4141 ggggagggga tctaatggcg ttggagtttg ttcacatttg gtgggtggag actagtcagg
     4201 ccagcctggc gctggaagtc attcttggaa tttgcccctt tgagtttgga gcgaggctaa
     4261 ttctcaagcc tcttagcggt tcaaaggtat tttctaaacc cgtttccagg tgttgtgaaa
     4321 gccaccgcta attcaaagca atccggagcc accatggtga gcaagggcga ggaggataac
     4381 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     4441 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     4501 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     4561 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     4621 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     4681 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     4741 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     4801 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     4861 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     4921 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     4981 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     5041 ggcatggacg agctgtacaa gtctggcagc ggaggctctg gaggcaaaga aaagatgagc
     5101 aaagatggta aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtgaggatcc
     5161 cgtacgccta ggagcaggtt tccccaatga cacaaaacgt gcaacttgaa actccgcctg
     5221 gtctttccag gtctagaggg gtaacacttt gtactgcgtt tggctccacg ctcgatccac
     5281 tggcgagtgt tagtaacagc actgttgctt cgtagcggag catgacggcc gtgggaactc
     5341 ctccttggta acaaggaccc acggggccaa aagccacgcc cacacgggcc cgtcatgtgt
     5401 gcaaccccag cacggcgact ttactgcgaa acccacttta aagtgacatt gaaactggta
     5461 cccacacact ggtgacaggc taaggatgcc cttcaggtac cccgaggtaa cacgcgacac
     5521 tcgggatctg agaaggggac tggggcttct ataaaagcgc tcggtttaaa aagcttctat
     5581 gcctgaatag gtgaccggag gtcggcacct ttcctttgca attactgacc ctatgaatac
     5641 actgactgtt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgactcac
     5701 tataggaggg ccaccatgaa gaaacctgaa ctgacagcaa cttctgttga gaagtttctc
     5761 attgaaaaat ttgattctgt ttctgatctc atgcagctgt ctgaaggtga agaaagcaga
     5821 gccttttctt ttgatgttgg aggaagaggt tatgttctga gggtcaattc ttgtgctgat
     5881 ggtttttaca aagacagata tgtttacaga cactttgcct ctgctgctct gccaattcca
     5941 gaagttctgg acattggaga attttctgaa tctctcacct actgcatcag cagaagagca
     6001 caaggagtca ctctccagga tctccctgaa actgagctgc cagctgttct gcaacctgtt
     6061 gctgaagcaa tggatgccat tgcagcagct gatctgagcc aaacctctgg atttggtcct
     6121 tttggtcccc aaggcattgg tcagtacacc acttggaggg atttcatttg tgccattgct
     6181 gatcctcatg tctatcactg gcagactgtg atggatgaca cagtttctgc ttctgttgct
     6241 caggcactgg atgaactcat gctgtgggca gaagattgtc ctgaagtcag acacctggtc
     6301 catgctgatt ttggaagcaa caatgttctg acagacaatg gcagaatcac tgcagtcatt
     6361 gactggtctg aagccatgtt tggagattct caatatgagg ttgccaacat ttttttttgg
     6421 agaccttggc tggcttgcat ggaacaacaa acaagatatt ttgaaagaag acacccagaa
     6481 ctggctggtt cccccagact gagagcctac atgctcagaa ttggcctgga ccaactgtat
     6541 caatctctgg ttgatggaaa ctttgatgat gctgcttggg cacaaggaag atgtgatgcc
     6601 attgtgaggt ctggtgctgg aactgttgga agaactcaaa ttgcaagaag gtctgctgct
     6661 gtttggactg atggatgtgt tgaagttctg gctgactctg gaaacaggag accctccaca
     6721 agacccagag ccaaggaatg aatattagct agattatccc taatacctgc caccccactc
     6781 ttaatcagtg gtggaagaac ggtctcagaa ctgtttgttt caattggcca tttaagttta
     6841 gtagtaaaag actggttaat gataacaatg catcgtaaaa ccttcagaag gaaaggagaa
     6901 tgttttgtgg accactttgg ttttcttttt tgcgtgtggc agttttaagt tattagtttt
     6961 taaaatcagt actttttaat ggaaacaact tgaccaaaaa tttgtcacag aattttgaga
     7021 cccattaaaa aagttaaatg agaaacctgt gtgttccttt ggtcaacacc gagacattta
     7081 ggtgaaagac atctaattct ggttttacga atctggaaac ttcttgaaaa tgtaattctt
     7141 gagttaacac ttctgggtgg agaatagggt tgttttcccc ccacataatt ggaaggggaa
     7201 ggaatatcat ttaaagctat gggagggttg ctttgattac aacactggag agaaatgcag
     7261 catgttgctg attgcctgtc actaaaacag gc
//
