LOCUS       sequence-366972-        7283 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Nucleus-targeted mCherry expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  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 7283)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-NLS
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        156..347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(179..199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        361..1667
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          688..1635
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     polyA_signal    1712..1833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1723..1742)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1777..1796
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2021..2609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2510..2529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2689..2992
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3107..4412
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3449..4380
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     regulatory      4419..4428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4425..5132
                     /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(4570..4588)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4838..4857
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5145..5165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5169..5189
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5193..5213
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5235..5679
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5712..5758
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5750..5768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5778..6803
                     /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(6815..7283,1..104)
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 ggagggttgc tttgattaca acactggaga gaaatgcagc atgttgctga ttgcctgtca
       61 ctaaaacagg ccaaaaactg agtccttggg ttgcatagaa agctgcctgc agggcctgaa
      121 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
      181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      241 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      301 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccact agtggagccg
      361 agagtaattc atacaaaagg agggatcgcc ttcgcaaggg gagagcccag ggaccgtccc
      421 taaattctca cagacccaaa tccctgtagc cgccccacga cagcgcgagg agcatgcgct
      481 cagggctgag cgcggggaga gcagagcaca caagctcata gaccctggtc gtggggggga
      541 ggaccgggga gctggcgcgg ggcaaactgg gaaagcggtg tcgtgtgctg gctccgccct
      601 cttcccgagg gtgggggaga acggtatata agtgcggcag tcgccttgga cgttcttttt
      661 cgcaacgggt ttgccgtcag aacgcaggtg aggggcgggt gtggcttccg cgggccgccg
      721 agctggaggt cctgctccga gcgggccggg ccccgctgtc gtcggcgggg attagctgcg
      781 agcattcccg cttcgagttg cgggcggcgc gggaggcaga gtgcgaggcc tagcggcaac
      841 cccgtagcct cgcctcgtgt ccggcttgag gcctagcgtg gtgtccgcgc cgccgccgcg
      901 tgctactccg gccgcactct ggtctttttt ttttttgttg ttgttgccct gctgccttcg
      961 attgccgttc agcaataggg gctaacaaag ggagggtgcg gggcttgctc gcccggagcc
     1021 cggagaggtc atggttgggg aggaatggag ggacaggagt ggcggctggg gcccgcccgc
     1081 cttcggagca catgtccgac gccacctgga tggggcgagg cctggggttt ttcccgaagc
     1141 aaccaggctg gggttagcgt gccgaggcca tgtggcccca gcacccggca cgatctggct
     1201 tggcggcgcc gcgttgccct gcctccctaa ctagggtgag gccatcccgt ccggcaccag
     1261 ttgcgtgcgt ggaaagatgg ccgctcccgg gccctgttgc aaggagctca aaatggagga
     1321 cgcggcagcc cggtggagcg ggcgggtgag tcacccacac aaaggaagag ggcctggtcc
     1381 ctcaccggct gctgcttcct gtgaccccgt ggtcctatcg gccgcaatag tcacctcggg
     1441 cttttgagca cggctagtcg cggcgggggg aggggatgta atggcgttgg agtttgttca
     1501 catttggtgg gtggagacta gtcaggccag cctggcgctg gaagtcattt ttggaatttg
     1561 tccccttgag ttttgagcgg agctaattct cgggcttctt agcggttcaa aggtatcttt
     1621 taaacccttt tttaggtgtt gtgaaaacca ccgctaattc aaagcaaccg gtgatatcaa
     1681 agatcttgta cagctagctg gccagacatg ataagataca ttgatgagtt tggacaaacc
     1741 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1801 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1861 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1921 ggtatggaaa tgttaattaa ctagccatga ccaaaatccc ttaacgtgag ttttcgttcc
     1981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2161 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2581 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2641 tggccttttg ctggcctttt gctcacatgt tcttaattaa cctgcaggcg ttacataact
     2701 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2761 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2821 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2881 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2941 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgatgatgcg
     3001 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     3061 ccaccccatt gacgtcaatg ggagtttgtt ttgactagtg gagccgagag taattcatac
     3121 aaaaggaggg atcgccttcg caaggggaga gcccagggac cgtccctaaa ttctcacaga
     3181 cccaaatccc tgtagccgcc ccacgacagc gcgaggagca tgcgcccagg gctgagcgcg
     3241 ggtagatcag agcacacaag ctcacagtcc ccggcggtgg ggggaggggc gcgctgagcg
     3301 ggggccaggg agctggcgcg gggcaaactg ggaaagtggt gtcgtgtgct ggctccgccc
     3361 tcttcccgag ggtgggggag aacggtatat aagtgcggta gtcgccttgg acgttctttt
     3421 tcgcaacggg tttgccgtca gaacgcaggt gagtggcggg tgtggcttcc gcgggccccg
     3481 gagctggagc cctgctctga gcgggccggg ctgatatgcg agtgtcgtcc gcagggttta
     3541 gctgtgagca ttcccacttc gagtggcggg cggtgcgggg gtgagagtgc gaggcctagc
     3601 ggcaaccccg tagcctcgcc tcgtgtccgg cttgaggcct agcgtggtgt ccgccgccgc
     3661 gtgccactcc ggccgcacta tgcgtttttt gtccttgctg ccctcgattg ccttccagca
     3721 gcatgggcta acaaagggag ggtgtggggc tcactcttaa ggagcccatg aagcttacgt
     3781 tggataggaa tggaagggca ggaggggcga ctggggcccg cccgccttcg gagcacatgt
     3841 ccgacgccac ctggatgggg cgaggcctgt ggctttccga agcaatcggg cgtgagttta
     3901 gcctacctgg gccatgtggc cctagcactg ggcacggtct ggcctggcgg tgccgcgttc
     3961 ccttgcctcc caacaagggt gaggccgtcc cgcccggcac cagttgcttg cgcggaaaga
     4021 tggccgctcc cggggccctg ttgcaaggag ctcaaaatgg aggacgcggc agcccggtgg
     4081 agcgggcggg tgagtcaccc acacaaagga agagggcctt gcccctcgcc ggccgctgct
     4141 tcctgtgacc ccgtggtcta tcggccgcat agtcacctcg ggcttctctt gagcaccgct
     4201 cgtcgcggcg gggggagggg atctaatggc gttggagttt gttcacattt ggtgggtgga
     4261 gactagtcag gccagcctgg cgctggaagt cattcttgga atttgcccct ttgagtttgg
     4321 agcgaggcta attctcaagc ctcttagcgg ttcaaaggta ttttctaaac ccgtttccag
     4381 gtgttgtgaa agccaccgct aattcaaagc aatccggagc caccatggtg agcaagggcg
     4441 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     4501 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     4561 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     4621 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     4681 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     4741 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     4801 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     4861 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     4921 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     4981 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     5041 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     5101 actccaccgg cggcatggac gagctgtaca agggaagctc cgatccaaaa aagaagagaa
     5161 aggtagaccc caaaaagaaa aggaaggtgg accctaagaa aaagcgcaaa gtgtgaggat
     5221 cccgtacgcc taggagcagg tttccccaat gacacaaaac gtgcaacttg aaactccgcc
     5281 tggtctttcc aggtctagag gggtaacact ttgtactgcg tttggctcca cgctcgatcc
     5341 actggcgagt gttagtaaca gcactgttgc ttcgtagcgg agcatgacgg ccgtgggaac
     5401 tcctccttgg taacaaggac ccacggggcc aaaagccacg cccacacggg cccgtcatgt
     5461 gtgcaacccc agcacggcga ctttactgcg aaacccactt taaagtgaca ttgaaactgg
     5521 tacccacaca ctggtgacag gctaaggatg cccttcaggt accccgaggt aacacgcgac
     5581 actcgggatc tgagaagggg actggggctt ctataaaagc gctcggttta aaaagcttct
     5641 atgcctgaat aggtgaccgg aggtcggcac ctttcctttg caattactga ccctatgaat
     5701 acactgactg tttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgactc
     5761 actataggag ggccaccatg aagaaacctg aactgacagc aacttctgtt gagaagtttc
     5821 tcattgaaaa atttgattct gtttctgatc tcatgcagct gtctgaaggt gaagaaagca
     5881 gagccttttc ttttgatgtt ggaggaagag gttatgttct gagggtcaat tcttgtgctg
     5941 atggttttta caaagacaga tatgtttaca gacactttgc ctctgctgct ctgccaattc
     6001 cagaagttct ggacattgga gaattttctg aatctctcac ctactgcatc agcagaagag
     6061 cacaaggagt cactctccag gatctccctg aaactgagct gccagctgtt ctgcaacctg
     6121 ttgctgaagc aatggatgcc attgcagcag ctgatctgag ccaaacctct ggatttggtc
     6181 cttttggtcc ccaaggcatt ggtcagtaca ccacttggag ggatttcatt tgtgccattg
     6241 ctgatcctca tgtctatcac tggcagactg tgatggatga cacagtttct gcttctgttg
     6301 ctcaggcact ggatgaactc atgctgtggg cagaagattg tcctgaagtc agacacctgg
     6361 tccatgctga ttttggaagc aacaatgttc tgacagacaa tggcagaatc actgcagtca
     6421 ttgactggtc tgaagccatg tttggagatt ctcaatatga ggttgccaac attttttttt
     6481 ggagaccttg gctggcttgc atggaacaac aaacaagata ttttgaaaga agacacccag
     6541 aactggctgg ttcccccaga ctgagagcct acatgctcag aattggcctg gaccaactgt
     6601 atcaatctct ggttgatgga aactttgatg atgctgcttg ggcacaagga agatgtgatg
     6661 ccattgtgag gtctggtgct ggaactgttg gaagaactca aattgcaaga aggtctgctg
     6721 ctgtttggac tgatggatgt gttgaagttc tggctgactc tggaaacagg agaccctcca
     6781 caagacccag agccaaggaa tgaatattag ctagattatc cctaatacct gccaccccac
     6841 tcttaatcag tggtggaaga acggtctcag aactgtttgt ttcaattggc catttaagtt
     6901 tagtagtaaa agactggtta atgataacaa tgcatcgtaa aaccttcaga aggaaaggag
     6961 aatgttttgt ggaccacttt ggttttcttt tttgcgtgtg gcagttttaa gttattagtt
     7021 tttaaaatca gtacttttta atggaaacaa cttgaccaaa aatttgtcac agaattttga
     7081 gacccattaa aaaagttaaa tgagaaacct gtgtgttcct ttggtcaaca ccgagacatt
     7141 taggtgaaag acatctaatt ctggttttac gaatctggaa acttcttgaa aatgtaattc
     7201 ttgagttaac acttctgggt ggagaatagg gttgttttcc ccccacataa ttggaagggg
     7261 aaggaatatc atttaaagct atg
//
