LOCUS       sequence-366974-        8984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear membrane-targeted mCherry expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8984)
  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 8984)
  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-LaminB1
FEATURES             Location/Qualifiers
     source          1..8984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..306
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(138..158)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        320..1626
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          647..1594
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1671..1792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1682..1701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1736..1755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1980..2568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2469..2488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2648..2951
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3066..4371
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3408..4339
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      4378..4387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4384..5091
                     /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(4529..4547)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4797..4816
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6895..7339
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        7372..7418
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7410..7428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7438..8463
                     /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(8475..8984,1..63)
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 tgttgctgat tgcctgtcac taaaacaggc caaaaactga gtccttgggt tgcatagaaa
       61 gctgcctgca gggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
      121 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
      181 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
      241 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
      301 agtcccacta gtggagccga gagtaattca tacaaaagga gggatcgcct tcgcaagggg
      361 agagcccagg gaccgtccct aaattctcac agacccaaat ccctgtagcc gccccacgac
      421 agcgcgagga gcatgcgctc agggctgagc gcggggagag cagagcacac aagctcatag
      481 accctggtcg tgggggggag gaccggggag ctggcgcggg gcaaactggg aaagcggtgt
      541 cgtgtgctgg ctccgccctc ttcccgaggg tgggggagaa cggtatataa gtgcggcagt
      601 cgccttggac gttctttttc gcaacgggtt tgccgtcaga acgcaggtga ggggcgggtg
      661 tggcttccgc gggccgccga gctggaggtc ctgctccgag cgggccgggc cccgctgtcg
      721 tcggcgggga ttagctgcga gcattcccgc ttcgagttgc gggcggcgcg ggaggcagag
      781 tgcgaggcct agcggcaacc ccgtagcctc gcctcgtgtc cggcttgagg cctagcgtgg
      841 tgtccgcgcc gccgccgcgt gctactccgg ccgcactctg gtcttttttt tttttgttgt
      901 tgttgccctg ctgccttcga ttgccgttca gcaatagggg ctaacaaagg gagggtgcgg
      961 ggcttgctcg cccggagccc ggagaggtca tggttgggga ggaatggagg gacaggagtg
     1021 gcggctgggg cccgcccgcc ttcggagcac atgtccgacg ccacctggat ggggcgaggc
     1081 ctggggtttt tcccgaagca accaggctgg ggttagcgtg ccgaggccat gtggccccag
     1141 cacccggcac gatctggctt ggcggcgccg cgttgccctg cctccctaac tagggtgagg
     1201 ccatcccgtc cggcaccagt tgcgtgcgtg gaaagatggc cgctcccggg ccctgttgca
     1261 aggagctcaa aatggaggac gcggcagccc ggtggagcgg gcgggtgagt cacccacaca
     1321 aaggaagagg gcctggtccc tcaccggctg ctgcttcctg tgaccccgtg gtcctatcgg
     1381 ccgcaatagt cacctcgggc ttttgagcac ggctagtcgc ggcgggggga ggggatgtaa
     1441 tggcgttgga gtttgttcac atttggtggg tggagactag tcaggccagc ctggcgctgg
     1501 aagtcatttt tggaatttgt ccccttgagt tttgagcgga gctaattctc gggcttctta
     1561 gcggttcaaa ggtatctttt aaaccctttt ttaggtgttg tgaaaaccac cgctaattca
     1621 aagcaaccgg tgatatcaaa gatcttgtac agctagctgg ccagacatga taagatacat
     1681 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1741 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1801 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1861 gtaaaacctc tacaaatgtg gtatggaaat gttaattaac tagccatgac caaaatccct
     1921 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1981 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2041 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2101 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2161 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2221 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2281 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2341 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2401 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2461 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2521 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2581 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt cttaattaac
     2641 ctgcaggcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2701 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2761 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2821 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     2881 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     2941 ctattaccat gatgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     3001 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tgactagtgg
     3061 agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag cccagggacc
     3121 gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg cgaggagcat
     3181 gcgcccaggg ctgagcgcgg gtagatcaga gcacacaagc tcacagtccc cggcggtggg
     3241 gggaggggcg cgctgagcgg gggccaggga gctggcgcgg ggcaaactgg gaaagtggtg
     3301 tcgtgtgctg gctccgccct cttcccgagg gtgggggaga acggtatata agtgcggtag
     3361 tcgccttgga cgttcttttt cgcaacgggt ttgccgtcag aacgcaggtg agtggcgggt
     3421 gtggcttccg cgggccccgg agctggagcc ctgctctgag cgggccgggc tgatatgcga
     3481 gtgtcgtccg cagggtttag ctgtgagcat tcccacttcg agtggcgggc ggtgcggggg
     3541 tgagagtgcg aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta
     3601 gcgtggtgtc cgccgccgcg tgccactccg gccgcactat gcgttttttg tccttgctgc
     3661 cctcgattgc cttccagcag catgggctaa caaagggagg gtgtggggct cactcttaag
     3721 gagcccatga agcttacgtt ggataggaat ggaagggcag gaggggcgac tggggcccgc
     3781 ccgccttcgg agcacatgtc cgacgccacc tggatggggc gaggcctgtg gctttccgaa
     3841 gcaatcgggc gtgagtttag cctacctggg ccatgtggcc ctagcactgg gcacggtctg
     3901 gcctggcggt gccgcgttcc cttgcctccc aacaagggtg aggccgtccc gcccggcacc
     3961 agttgcttgc gcggaaagat ggccgctccc ggggccctgt tgcaaggagc tcaaaatgga
     4021 ggacgcggca gcccggtgga gcgggcgggt gagtcaccca cacaaaggaa gagggccttg
     4081 cccctcgccg gccgctgctt cctgtgaccc cgtggtctat cggccgcata gtcacctcgg
     4141 gcttctcttg agcaccgctc gtcgcggcgg ggggagggga tctaatggcg ttggagtttg
     4201 ttcacatttg gtgggtggag actagtcagg ccagcctggc gctggaagtc attcttggaa
     4261 tttgcccctt tgagtttgga gcgaggctaa ttctcaagcc tcttagcggt tcaaaggtat
     4321 tttctaaacc cgtttccagg tgttgtgaaa gccaccgcta attcaaagca atccggagcc
     4381 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     4441 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     4501 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     4561 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     4621 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     4681 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4741 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4801 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4861 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4921 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4981 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     5041 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtctggcagc
     5101 ggaggctctg gaggcatggc gactgcgacc cccgtgccgc cgcggatggg cagccgcgct
     5161 ggcggcccca ccacgccgct gagccccacg cgcctgtcgc ggctccagga gaaggaggag
     5221 ctgcgcgagc tcaatgaccg gctggcggtg tacatcgaca aggtgcgcag cctggagacg
     5281 gagaacagcg cgctgcagct gcaggtgacg gagcgcgagg aggtgcgcgg ccgtgagctc
     5341 accggcctca aggcgctcta cgagaccgag ctggccgacg cgcgacgcgc gctcgacgac
     5401 acggcccgcg agcgcgccaa gctgcagatc gagctgggca agtgcaaggc ggaacacgac
     5461 cagctgctcc tcaactatgc taagaaggaa tctgatctta atggcgccca gatcaagctt
     5521 cgagaatatg aagcagcact caattcgaaa gatgcagctc ttgctactgc acttggtgac
     5581 aaaaaaagtt tagagggaga tttggaggat ctgaaggatc agattgccca gttggaagcc
     5641 tccttagctg cagccaaaaa acagttagca gatgaaactt tacttaaagt agatttggag
     5701 aatcgttgtc agagccttac tgaggacttg gagtttcgca aaagcatgta tgaagaggag
     5761 attaacgaga ccagaaggaa gcatgaaacg cgcttggtag aggtggattc tgggcgtcaa
     5821 attgagtatg agtacaagct ggcgcaagcc cttcatgaga tgagagagca acatgatgcc
     5881 caagtgaggc tgtataagga ggagctggag cagacttacc atgccaaact tgagaatgcc
     5941 agactgtcat cagagatgaa tacttctact gtcaacagtg ccagggaaga actgatggaa
     6001 agccgcatga gaattgagag cctttcatcc cagctttcta atctacagaa agagtctaga
     6061 gcatgtttgg aaaggattca agaattagag gacttgcttg ctaaagaaaa agacaactct
     6121 cgtcgcatgc tgacagacaa agagagagag atggcggaaa taagggatca aatgcagcaa
     6181 cagctgaatg actatgaaca gcttcttgat gtaaagttag ccctggacat ggaaatcagt
     6241 gcttacagga aactcttaga aggcgaagaa gagaggttga agctgtctcc aagcccttct
     6301 tcccgtgtga cagtatcccg agcatcctca agtcgtagtg tacgtacaac tagaggaaag
     6361 cggaagaggg ttgatgtgga agaatcagag gcgagtagta gtgttagcat ctctcattcc
     6421 gcctcagcca ctggaaatgt ttgcatcgaa gaaattgatg ttgatgggaa atttatccgc
     6481 ttgaagaaca cttctgaaca ggatcaacca atgggaggct gggagatgat cagaaaaatt
     6541 ggagacacat cagtcagtta taaatatacc tcaagatatg tgctgaaggc aggccagact
     6601 gttacaattt gggctgcaaa cgctggtgtc acagccagcc ccccaactga cctcatctgg
     6661 aagaaccaga actcgtgggg cactggcgaa gatgtgaagg ttatattgaa aaattctcag
     6721 ggagaggagg ttgctcaaag aagtacagtc tttaaaacaa ccatacctga agaagaggag
     6781 gaggaggaag aagcagctgg agtggttgtt gaggaagaac ttttccacca gcagggaacc
     6841 ccaagagcat ccaatagaag ctgtgcaatt atgtgaggat cccgtacgcc taggagcagg
     6901 tttccccaat gacacaaaac gtgcaacttg aaactccgcc tggtctttcc aggtctagag
     6961 gggtaacact ttgtactgcg tttggctcca cgctcgatcc actggcgagt gttagtaaca
     7021 gcactgttgc ttcgtagcgg agcatgacgg ccgtgggaac tcctccttgg taacaaggac
     7081 ccacggggcc aaaagccacg cccacacggg cccgtcatgt gtgcaacccc agcacggcga
     7141 ctttactgcg aaacccactt taaagtgaca ttgaaactgg tacccacaca ctggtgacag
     7201 gctaaggatg cccttcaggt accccgaggt aacacgcgac actcgggatc tgagaagggg
     7261 actggggctt ctataaaagc gctcggttta aaaagcttct atgcctgaat aggtgaccgg
     7321 aggtcggcac ctttcctttg caattactga ccctatgaat acactgactg tttgacaatt
     7381 aatcatcggc atagtatatc ggcatagtat aatacgactc actataggag ggccaccatg
     7441 aagaaacctg aactgacagc aacttctgtt gagaagtttc tcattgaaaa atttgattct
     7501 gtttctgatc tcatgcagct gtctgaaggt gaagaaagca gagccttttc ttttgatgtt
     7561 ggaggaagag gttatgttct gagggtcaat tcttgtgctg atggttttta caaagacaga
     7621 tatgtttaca gacactttgc ctctgctgct ctgccaattc cagaagttct ggacattgga
     7681 gaattttctg aatctctcac ctactgcatc agcagaagag cacaaggagt cactctccag
     7741 gatctccctg aaactgagct gccagctgtt ctgcaacctg ttgctgaagc aatggatgcc
     7801 attgcagcag ctgatctgag ccaaacctct ggatttggtc cttttggtcc ccaaggcatt
     7861 ggtcagtaca ccacttggag ggatttcatt tgtgccattg ctgatcctca tgtctatcac
     7921 tggcagactg tgatggatga cacagtttct gcttctgttg ctcaggcact ggatgaactc
     7981 atgctgtggg cagaagattg tcctgaagtc agacacctgg tccatgctga ttttggaagc
     8041 aacaatgttc tgacagacaa tggcagaatc actgcagtca ttgactggtc tgaagccatg
     8101 tttggagatt ctcaatatga ggttgccaac attttttttt ggagaccttg gctggcttgc
     8161 atggaacaac aaacaagata ttttgaaaga agacacccag aactggctgg ttcccccaga
     8221 ctgagagcct acatgctcag aattggcctg gaccaactgt atcaatctct ggttgatgga
     8281 aactttgatg atgctgcttg ggcacaagga agatgtgatg ccattgtgag gtctggtgct
     8341 ggaactgttg gaagaactca aattgcaaga aggtctgctg ctgtttggac tgatggatgt
     8401 gttgaagttc tggctgactc tggaaacagg agaccctcca caagacccag agccaaggaa
     8461 tgaatattag ctagattatc cctaatacct gccaccccac tcttaatcag tggtggaaga
     8521 acggtctcag aactgtttgt ttcaattggc catttaagtt tagtagtaaa agactggtta
     8581 atgataacaa tgcatcgtaa aaccttcaga aggaaaggag aatgttttgt ggaccacttt
     8641 ggttttcttt tttgcgtgtg gcagttttaa gttattagtt tttaaaatca gtacttttta
     8701 atggaaacaa cttgaccaaa aatttgtcac agaattttga gacccattaa aaaagttaaa
     8761 tgagaaacct gtgtgttcct ttggtcaaca ccgagacatt taggtgaaag acatctaatt
     8821 ctggttttac gaatctggaa acttcttgaa aatgtaattc ttgagttaac acttctgggt
     8881 ggagaatagg gttgttttcc ccccacataa ttggaagggg aaggaatatc atttaaagct
     8941 atgggagggt tgctttgatt acaacactgg agagaaatgc agca
//
