LOCUS       sequence-351691-        5067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a protein of interest fused to the 
            C-terminus of FKBP-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  AUTHORS   Yoshikawa M, Yoshii T, Ikuta M, Tsukiji S
  TITLE     Synthetic Protein Condensates That Inducibly Recruit and Release 
            Protein Activity in Living Cells.
  JOURNAL   J Am Chem Soc. 2021 May 5;143(17):6434-6446. doi: 
            10.1021/jacs.0c12375. Epub 2021 Apr 23.
  PUBMED    33890764
REFERENCE   2  (bases 1 to 5067)
  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: 
            "10.1021/jacs.0c12375"; journalName: "J Am Chem Soc"; date: 
            "2021-05-5- 5"; volume: "143"; issue: "17"; pages: "6434-6446"
COMMENT     Sequence Label: pCMV-FKBP-mCherry-MCS
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      589..598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(595..597,598..918)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      943..952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             949..1656
                     /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(1094..1112)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1362..1381
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1657..1722
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1846..1967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1883..1902)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1937..1956
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1974..2429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2111..2132)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2323..2342
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2456..2560
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2562..2919
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2562..2753
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2585..2605)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2770..2905
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2954..3748
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2954..3748
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3008..3027)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3618..3637
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3936..3955)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3980..4027
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4356..4944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4845..4864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagccgc caccatggga
      601 gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc
      661 tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac
      721 agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa
      781 ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc
      841 tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg
      901 gagcttctaa aactggaagg ctccggtgcc agtgcaccgg tcgccaccat ggtgagcaag
      961 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     1021 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1081 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     1141 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1201 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1261 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1321 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1381 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1441 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     1501 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     1561 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     1621 cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcaa
     1681 gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg
     1741 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1801 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1861 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1921 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     1981 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2041 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2101 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2161 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2221 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2281 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2341 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2401 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2461 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2521 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2581 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2641 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2761 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2821 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2881 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2941 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3001 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3061 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3121 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3181 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3241 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3301 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3361 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3421 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3481 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3541 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3601 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3661 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3721 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3781 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3841 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3901 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3961 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4021 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4081 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4141 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4501 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtatc accgcctaca
     4561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4741 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4981 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5041 cgtattaccg ccatgcatta gttatta
//
