LOCUS       sequence-351707-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SOS1(Rem-Cdc25) fused to FKBP-mTagBFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  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 6498)
  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-mTagBFP2-SOScat
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        375..578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     529..549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(614..616,617..937)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      962..971
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             968..1678
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    3296..3417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3333..3352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3387..3406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3424..3879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3773..3792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3906..4010
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4012..4369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4012..4203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4035..4055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4220..4355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4404..5198
                     /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             4404..5198
                     /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(4458..4477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5068..5087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5386..5405)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5430..5477
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5806..6394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6295..6314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      601 gctagccgcc accatgggag tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt
      661 ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa
      721 atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt
      781 gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac
      841 tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc
      901 cactctcgtc ttcgatgtgg agcttctaaa actggaaggc tccggtgcca gtgcaccggt
      961 cgccaccatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta
     1021 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc
     1081 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
     1141 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca
     1201 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac
     1261 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
     1321 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca
     1381 gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct
     1441 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc
     1501 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta
     1561 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca
     1621 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatag
     1681 atctcgagct caagcttcgc agatgaggct gcctagtgct gatgtttata gatttgcaga
     1741 gcctgactct gaagagaata ttatatttga agagaacatg cagcccaagg ctggaattcc
     1801 aattatcaaa gcaggaactg ttattaaact tatagagagg cttacgtacc atatgtacgc
     1861 agatcccaat tttgttcgga catttcttac aacatacaga tccttttgca aacctcaaga
     1921 actactgagt cttataatag aaaggtttga aattccagag cctgagccaa cagaagctga
     1981 tcgcatagct atagagaatg gagatcaacc cttgagtgca gaactgaaaa gatttagaaa
     2041 agaatatata cagcctgtgc aactgcgagt attaaatgta tgtcggcact gggtagagca
     2101 ccacttctat gattttgaaa gagatgcata tcttttgcaa cgaatggaag aatttattgg
     2161 aacagtaaga ggtaaagcaa tgaaaaaatg ggttgaatcc atcactaaaa taatccaaag
     2221 gaaaaaaatt gcaagagaca atggaccagg tcataatatt acatttcaga gttcacctcc
     2281 cacagttgag tggcatataa gcagacctgg gcacatagag acttttgacc tgctcacctt
     2341 acacccaata gaaattgctc gacaactcac tttacttgaa tcagatctat accgagctgt
     2401 acagccatca gaattagttg gaagtgtgtg gacaaaagaa gacaaagaaa ttaactctcc
     2461 taatcttctg aaaatgattc gacataccac caacctcact ctgtggtttg agaaatgtat
     2521 tgtagaaact gaaaatttag aagaaagagt agctgtggtg agtcgaatta ttgagattct
     2581 acaagtcttt caagagttga acaactttaa tggtgtcctt gaggttgtca gtgctatgaa
     2641 ttcatcacct gtttacagac tagaccacac atttgagcaa ataccaagtc gccagaagaa
     2701 aattttagaa gaagctcatg aattgagtga agatcactat aagaaatatt tggcaaaact
     2761 caggtctatt aatccaccat gtgtgccttt ctttggaatt tatctcacta atatcttgaa
     2821 aacagaagaa ggcaaccctg aggtcctaaa aagacatgga aaagagctta taaactttag
     2881 caaaaggagg aaagtagcag aaataacagg agagatccag cagtaccaaa atcagcctta
     2941 ctgtttacga gtagaatcag atatcaaaag gttctttgaa aacttgaatc cgatgggaaa
     3001 tagcatggag aaggaattta cagattatct tttcaacaaa tccctagaaa tagaaccacg
     3061 aaaccctaag cctctcccaa gatttccaaa aaaatatagc tatcccctaa aatctcctgg
     3121 tgttcgtcca tcaaacccaa gaccaggtac ctaaaccgcg ggcccgggat ccaccggatc
     3181 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3241 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3301 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3361 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3421 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3481 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3541 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3601 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3661 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3721 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3781 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3841 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4021 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4081 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4201 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4261 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4321 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4381 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4441 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4501 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4561 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4621 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4681 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4741 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4801 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4861 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4921 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4981 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5041 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5101 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5161 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5221 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5341 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5401 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5461 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5521 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5581 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5641 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5701 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5761 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5821 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5881 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5941 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtatc
     6001 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6061 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6121 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6181 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6241 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6301 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6361 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6421 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6481 tgtggataac cgtattac
//
