LOCUS       sequence-351689-        6207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Vav2(DH-PH-CRD) fused to FKBP-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  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 6207)
  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-Vav2cat
FEATURES             Location/Qualifiers
     source          1..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        351..554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     505..525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      584..593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(590..592,593..913)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      938..947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             944..1651
                     /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(1089..1107)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1357..1376
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2981..3102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3018..3037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3072..3091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3109..3564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3246..3267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3458..3477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3591..3695
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3697..4054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3697..3888
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3720..3740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3905..4040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4089..4883
                     /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             4089..4883
                     /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(4143..4162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4753..4772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5071..5090)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5115..5162
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5491..6079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5980..5999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
       61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      541 atataagcag agctggttta gtgaaccgtc agatccgcta gccgccacca tgggagtgca
      601 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt
      661 ggtgcactac accgggatgc ttgaagatgg aaagaaattt gattcctccc gggacagaaa
      721 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
      781 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
      841 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
      901 tctaaaactg gaaggctccg gtgccagtgc accggtcgcc accatggtga gcaagggcga
      961 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     1021 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1081 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1141 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1201 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1261 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1321 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1381 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1441 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1501 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1561 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1621 ctccaccggc ggcatggacg agctgtacaa gtccggactc agatctcgag ggcccatgaa
     1681 aatgggcatg actgaggacg acaagagaag ctgctgcttg ttagagattc aggagaccga
     1741 ggccaagtac taccgcaccc tggaggacat tgagaagaac tacatgggtc ccttgcggct
     1801 ggtgctgagc ccggcggata tggctgctgt cttcatcaac ctggaggacc tcatcaaggt
     1861 gcatcacagc tttctgcgag ccatcgatgt gtccatgatg gctggtggca gtaccctggc
     1921 taaggtcttt ctggagttta aggaaaggct cctgatctat ggagagtact gtagccacat
     1981 ggaacacgct cagagtacac tgaaccagct cctcgccagc cgagaggact tcaggcagaa
     2041 agtggaggag tgcacactca gggttcagga tggcaagttc aagctgcaag acctgctggt
     2101 ggtgcccatg caacgggtgc tgaagtacca cctgctgctc aaggagctcc tgagccattc
     2161 tgcagaccga ccagaaagac aacagctcaa agaagccctg gaagccatgc aggacttggc
     2221 catgtacatc aatgaagtga agcgggacaa ggagaccttg aagaagatta gcgagttcca
     2281 gtgctccata gaaaacctgc aagtgaagct ggaggaattt gggaggccaa agattgacgg
     2341 ggagcttaaa gtccggtcca tagtcaacca caccaagcaa gacaggtacc tgttcctatt
     2401 tgacaaggtg gtcatcgtgt gcaagaggaa gggctacagc tatgagctga aggaggtcat
     2461 tgagctgctc ttccacaaga tgaccgatga cccgatgcac aacaaggaca tcaagaagtg
     2521 gtcctatggc ttctacctga ttcacctcca aggaaagcaa ggctttcagt tcttctgcaa
     2581 gacggaagac atgaagcgga agtggatgga gcagttcgag atggccatgt caaacatcaa
     2641 gccagataag gccaatgcca accatcatag cttccagatg tacacattcg acaagactac
     2701 caactgcaaa gcctgcaaga tgtttctcag gggtaccttc taccagggat acctgtgtac
     2761 cagatgtggc gtcggggcac acaaggaatg cctggaggtg atccccccct gcaagtaacc
     2821 cgggatactc gcggccgcca ccgcgggccc gggatccacc ggatctagat aactgatcat
     2881 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2941 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3061 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3121 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3181 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3241 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3301 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3361 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3421 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3481 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3541 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3601 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3661 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3961 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4021 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4081 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4141 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4201 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4261 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4321 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4381 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4441 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4501 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4561 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4621 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4681 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4741 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4801 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4861 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4921 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4981 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5041 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5101 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5161 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5221 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5281 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5341 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5401 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5461 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5521 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5581 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5641 tctagtgtag ccgtagttag gccaccactt caagaactct gtatcaccgc ctacatacct
     5701 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5761 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5821 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5881 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5941 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6001 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6061 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6121 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6181 taccgccatg cattagttat taatagt
//
