LOCUS       sequence-366975-        7241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic mCherry expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7241)
  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 7241)
  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-NES
FEATURES             Location/Qualifiers
     source          1..7241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..285
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(117..137)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        299..1605
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          626..1573
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1650..1771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1661..1680)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1715..1734
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1959..2547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2448..2467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2627..2930
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3045..4350
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3387..4318
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      4357..4366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4363..5070
                     /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(4508..4526)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4776..4795
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5131..5575
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5608..5654
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5646..5664
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5674..6699
                     /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(6711..7241,1..42)
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 aaaacaggcc aaaaactgag tccttgggtt gcatagaaag ctgcctgcag ggcctgaaat
       61 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
      121 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      181 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
      241 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccactag tggagccgag
      301 agtaattcat acaaaaggag ggatcgcctt cgcaagggga gagcccaggg accgtcccta
      361 aattctcaca gacccaaatc cctgtagccg ccccacgaca gcgcgaggag catgcgctca
      421 gggctgagcg cggggagagc agagcacaca agctcataga ccctggtcgt gggggggagg
      481 accggggagc tggcgcgggg caaactggga aagcggtgtc gtgtgctggc tccgccctct
      541 tcccgagggt gggggagaac ggtatataag tgcggcagtc gccttggacg ttctttttcg
      601 caacgggttt gccgtcagaa cgcaggtgag gggcgggtgt ggcttccgcg ggccgccgag
      661 ctggaggtcc tgctccgagc gggccgggcc ccgctgtcgt cggcggggat tagctgcgag
      721 cattcccgct tcgagttgcg ggcggcgcgg gaggcagagt gcgaggccta gcggcaaccc
      781 cgtagcctcg cctcgtgtcc ggcttgaggc ctagcgtggt gtccgcgccg ccgccgcgtg
      841 ctactccggc cgcactctgg tctttttttt ttttgttgtt gttgccctgc tgccttcgat
      901 tgccgttcag caataggggc taacaaaggg agggtgcggg gcttgctcgc ccggagcccg
      961 gagaggtcat ggttggggag gaatggaggg acaggagtgg cggctggggc ccgcccgcct
     1021 tcggagcaca tgtccgacgc cacctggatg gggcgaggcc tggggttttt cccgaagcaa
     1081 ccaggctggg gttagcgtgc cgaggccatg tggccccagc acccggcacg atctggcttg
     1141 gcggcgccgc gttgccctgc ctccctaact agggtgaggc catcccgtcc ggcaccagtt
     1201 gcgtgcgtgg aaagatggcc gctcccgggc cctgttgcaa ggagctcaaa atggaggacg
     1261 cggcagcccg gtggagcggg cgggtgagtc acccacacaa aggaagaggg cctggtccct
     1321 caccggctgc tgcttcctgt gaccccgtgg tcctatcggc cgcaatagtc acctcgggct
     1381 tttgagcacg gctagtcgcg gcggggggag gggatgtaat ggcgttggag tttgttcaca
     1441 tttggtgggt ggagactagt caggccagcc tggcgctgga agtcattttt ggaatttgtc
     1501 cccttgagtt ttgagcggag ctaattctcg ggcttcttag cggttcaaag gtatctttta
     1561 aacccttttt taggtgttgt gaaaaccacc gctaattcaa agcaaccggt gatatcaaag
     1621 atcttgtaca gctagctggc cagacatgat aagatacatt gatgagtttg gacaaaccac
     1681 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1741 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1801 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1861 tatggaaatg ttaattaact agccatgacc aaaatccctt aacgtgagtt ttcgttccac
     1921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2581 gccttttgct ggccttttgc tcacatgttc ttaattaacc tgcaggcgtt acataactta
     2641 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2701 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2761 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2821 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2881 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg atgatgcggt
     2941 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     3001 accccattga cgtcaatggg agtttgtttt gactagtgga gccgagagta attcatacaa
     3061 aaggagggat cgccttcgca aggggagagc ccagggaccg tccctaaatt ctcacagacc
     3121 caaatccctg tagccgcccc acgacagcgc gaggagcatg cgcccagggc tgagcgcggg
     3181 tagatcagag cacacaagct cacagtcccc ggcggtgggg ggaggggcgc gctgagcggg
     3241 ggccagggag ctggcgcggg gcaaactggg aaagtggtgt cgtgtgctgg ctccgccctc
     3301 ttcccgaggg tgggggagaa cggtatataa gtgcggtagt cgccttggac gttctttttc
     3361 gcaacgggtt tgccgtcaga acgcaggtga gtggcgggtg tggcttccgc gggccccgga
     3421 gctggagccc tgctctgagc gggccgggct gatatgcgag tgtcgtccgc agggtttagc
     3481 tgtgagcatt cccacttcga gtggcgggcg gtgcgggggt gagagtgcga ggcctagcgg
     3541 caaccccgta gcctcgcctc gtgtccggct tgaggcctag cgtggtgtcc gccgccgcgt
     3601 gccactccgg ccgcactatg cgttttttgt ccttgctgcc ctcgattgcc ttccagcagc
     3661 atgggctaac aaagggaggg tgtggggctc actcttaagg agcccatgaa gcttacgttg
     3721 gataggaatg gaagggcagg aggggcgact ggggcccgcc cgccttcgga gcacatgtcc
     3781 gacgccacct ggatggggcg aggcctgtgg ctttccgaag caatcgggcg tgagtttagc
     3841 ctacctgggc catgtggccc tagcactggg cacggtctgg cctggcggtg ccgcgttccc
     3901 ttgcctccca acaagggtga ggccgtcccg cccggcacca gttgcttgcg cggaaagatg
     3961 gccgctcccg gggccctgtt gcaaggagct caaaatggag gacgcggcag cccggtggag
     4021 cgggcgggtg agtcacccac acaaaggaag agggccttgc ccctcgccgg ccgctgcttc
     4081 ctgtgacccc gtggtctatc ggccgcatag tcacctcggg cttctcttga gcaccgctcg
     4141 tcgcggcggg gggaggggat ctaatggcgt tggagtttgt tcacatttgg tgggtggaga
     4201 ctagtcaggc cagcctggcg ctggaagtca ttcttggaat ttgccccttt gagtttggag
     4261 cgaggctaat tctcaagcct cttagcggtt caaaggtatt ttctaaaccc gtttccaggt
     4321 gttgtgaaag ccaccgctaa ttcaaagcaa tccggagcca ccatggtgag caagggcgag
     4381 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     4441 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     4501 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     4561 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     4621 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     4681 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     4741 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     4801 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     4861 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     4921 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     4981 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     5041 tccaccggcg gcatggacga gctgtacaaa ggaagctccc tccagaagaa actggaggaa
     5101 ctggaactgt gaggatcccg tacgcctagg agcaggtttc cccaatgaca caaaacgtgc
     5161 aacttgaaac tccgcctggt ctttccaggt ctagaggggt aacactttgt actgcgtttg
     5221 gctccacgct cgatccactg gcgagtgtta gtaacagcac tgttgcttcg tagcggagca
     5281 tgacggccgt gggaactcct ccttggtaac aaggacccac ggggccaaaa gccacgccca
     5341 cacgggcccg tcatgtgtgc aaccccagca cggcgacttt actgcgaaac ccactttaaa
     5401 gtgacattga aactggtacc cacacactgg tgacaggcta aggatgccct tcaggtaccc
     5461 cgaggtaaca cgcgacactc gggatctgag aaggggactg gggcttctat aaaagcgctc
     5521 ggtttaaaaa gcttctatgc ctgaataggt gaccggaggt cggcaccttt cctttgcaat
     5581 tactgaccct atgaatacac tgactgtttg acaattaatc atcggcatag tatatcggca
     5641 tagtataata cgactcacta taggagggcc accatgaaga aacctgaact gacagcaact
     5701 tctgttgaga agtttctcat tgaaaaattt gattctgttt ctgatctcat gcagctgtct
     5761 gaaggtgaag aaagcagagc cttttctttt gatgttggag gaagaggtta tgttctgagg
     5821 gtcaattctt gtgctgatgg tttttacaaa gacagatatg tttacagaca ctttgcctct
     5881 gctgctctgc caattccaga agttctggac attggagaat tttctgaatc tctcacctac
     5941 tgcatcagca gaagagcaca aggagtcact ctccaggatc tccctgaaac tgagctgcca
     6001 gctgttctgc aacctgttgc tgaagcaatg gatgccattg cagcagctga tctgagccaa
     6061 acctctggat ttggtccttt tggtccccaa ggcattggtc agtacaccac ttggagggat
     6121 ttcatttgtg ccattgctga tcctcatgtc tatcactggc agactgtgat ggatgacaca
     6181 gtttctgctt ctgttgctca ggcactggat gaactcatgc tgtgggcaga agattgtcct
     6241 gaagtcagac acctggtcca tgctgatttt ggaagcaaca atgttctgac agacaatggc
     6301 agaatcactg cagtcattga ctggtctgaa gccatgtttg gagattctca atatgaggtt
     6361 gccaacattt ttttttggag accttggctg gcttgcatgg aacaacaaac aagatatttt
     6421 gaaagaagac acccagaact ggctggttcc cccagactga gagcctacat gctcagaatt
     6481 ggcctggacc aactgtatca atctctggtt gatggaaact ttgatgatgc tgcttgggca
     6541 caaggaagat gtgatgccat tgtgaggtct ggtgctggaa ctgttggaag aactcaaatt
     6601 gcaagaaggt ctgctgctgt ttggactgat ggatgtgttg aagttctggc tgactctgga
     6661 aacaggagac cctccacaag acccagagcc aaggaatgaa tattagctag attatcccta
     6721 atacctgcca ccccactctt aatcagtggt ggaagaacgg tctcagaact gtttgtttca
     6781 attggccatt taagtttagt agtaaaagac tggttaatga taacaatgca tcgtaaaacc
     6841 ttcagaagga aaggagaatg ttttgtggac cactttggtt ttcttttttg cgtgtggcag
     6901 ttttaagtta ttagttttta aaatcagtac tttttaatgg aaacaacttg accaaaaatt
     6961 tgtcacagaa ttttgagacc cattaaaaaa gttaaatgag aaacctgtgt gttcctttgg
     7021 tcaacaccga gacatttagg tgaaagacat ctaattctgg ttttacgaat ctggaaactt
     7081 cttgaaaatg taattcttga gttaacactt ctgggtggag aatagggttg ttttcccccc
     7141 acataattgg aaggggaagg aatatcattt aaagctatgg gagggttgct ttgattacaa
     7201 cactggagag aaatgcagca tgttgctgat tgcctgtcac t
//
