LOCUS       sequence-355798-        7957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SOS1(Rem-Cdc25) fused to 
            PB1-AzamiGreen-FRB-TEVcs-mTagBFP2-HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7957)
  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 7957)
  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-PB1-AG-FRB-TEVcs-mTagBFP2-HA-SOScat
FEATURES             Location/Qualifiers
     source          1..7957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      540..549
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1230..1907
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGVNFPPNGPVMQKKTLKWEPSTEKMYVRDGVLKGDVNMALL
                     LEGGGHYRCDFKTTYKAKKDVRLPDYHFVDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             1959..2237
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             2343..3053
                     /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"
     CDS             3066..3092
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4686..4807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4723..4742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4777..4796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4814..5269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4951..4972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5163..5182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5296..5400
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5402..5759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5402..5593
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5425..5445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5610..5745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5793..6587
                     /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             5793..6587
                     /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(5847..5866)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6457..6476
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6775..6794)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6819..6866
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7195..7783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7684..7703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcattg
      541 ccaccatggc gtcgctcacc gtgaaggcct accttctggg caaggaggac gcggcgcgcg
      601 agattcgccg cttcagcttc tgttgcagcc ccgagcctga ggcggaagcc gaggctgcgg
      661 cgggtccggg accctgcgag cggctgctga gccgggtggc cgccctgttc cccgcgctgc
      721 ggcctggcgg cttccaggcg cactaccgcg atgaggacgg ggacttggtt gccttttcca
      781 gtgacgagga attgacaatg gccatgtcct acgtgaagga tgacatcttc cgaatctaca
      841 ttaaagagaa aaccggttct gggagtggcg gtagtggtgc tggaggcagc gcaggctctg
      901 gagcaggcgg cagtgccgga agtggcgctg gagggtctgc cggatctgga gccggtggtt
      961 ctgccggctc cggcgctggt gggagcgctt ccggaagtag tcccggttct gggagtggcg
     1021 gtagtggtgc tggaggcagc gcaggctctg gagcaggcgg cagtgccgga agtggcgctg
     1081 gagggtctgc cggatctgga gccggtggtt ctgccggctc cggcgctggt gggagcgctt
     1141 ccggaagtag tcccggtaat tccgctgacg gcggcggagg atcgggtggt agtggtggtt
     1201 caggaggagg atcgacccaa ggaggatcca tggtgagcgt gatcaagccc gagatgaaga
     1261 tcaagctgtg catgaggggc accgtgaacg gccacaactt cgtgatcgag ggcgagggca
     1321 agggcaaccc ctacgagggc acccagatcc tggacctgaa cgtgaccgag ggcgcccccc
     1381 tgcccttcgc ctacgacatc ctgaccaccg tgttccagta cggcaacagg gccttcacca
     1441 agtaccccgc cgacatccag gactacttca agcagacctt ccccgagggc taccactggg
     1501 agaggagcat gacctacgag gaccagggca tctgcaccgc caccagcaac atcagcatga
     1561 ggggcgactg cttcttctac gacatcaggt tcgacggcgt gaacttcccc cccaacggcc
     1621 ccgtgatgca gaagaagacc ctgaagtggg agcccagcac cgagaagatg tacgtgaggg
     1681 acggcgtgct gaagggcgac gtgaacatgg ccctgctgct ggagggcggc ggccactaca
     1741 ggtgcgactt caagaccacc tacaaggcca agaaggacgt gaggctgccc gactaccact
     1801 tcgtggacca caggatcgag atcctgaagc acgacaagga ctacaacaag gtgaagctgt
     1861 acgagaacgc cgtggccagg tacagcatgc tgcccagcca ggccaagggt accggaactg
     1921 cagcagagaa ttcgggaaac tcgagaacaa agcttatgat cctctggcat gagatgtggc
     1981 atgaaggcct ggaagaggca tctcgtttgt actttgggga aaggaacgtg aaaggcatgt
     2041 ttgaggtgct ggagcccttg catgctatga tggaacgggg cccccagact ctgaaggaaa
     2101 catcctttaa tcaggcctat ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt
     2161 acatgaaatc agggaatgtc aaggacctcc tccaagcctg ggacctctat tatcatgtgt
     2221 tccgacgaat ctcaaagccc gggggtagtg gtgctggctc tggtgctggt agtggcgctg
     2281 gtgaaaacct gtattttcag ctggtcgatg gtggtagtgc tggtggttcc ggaggtgtcg
     2341 acatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg
     2401 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
     2461 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     2521 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
     2581 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     2641 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     2701 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     2761 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     2821 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacgccaaga
     2881 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     2941 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     3001 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt aatagatccg
     3061 gctcttaccc atacgatgtt ccagattacg ctagatctca gatgaggctg cctagtgctg
     3121 atgtttatag atttgcagag cctgactctg aagagaatat tatatttgaa gagaacatgc
     3181 agcccaaggc tggaattcca attatcaaag caggaactgt tattaaactt atagagaggc
     3241 ttacgtacca tatgtacgca gatcccaatt ttgttcggac atttcttaca acatacagat
     3301 ccttttgcaa acctcaagaa ctactgagtc ttataataga aaggtttgaa attccagagc
     3361 ctgagccaac agaagctgat cgcatagcta tagagaatgg agatcaaccc ttgagtgcag
     3421 aactgaaaag atttagaaaa gaatatatac agcctgtgca actgcgagta ttaaatgtat
     3481 gtcggcactg ggtagagcac cacttctatg attttgaaag agatgcatat cttttgcaac
     3541 gaatggaaga atttattgga acagtaagag gtaaagcaat gaaaaaatgg gttgaatcca
     3601 tcactaaaat aatccaaagg aaaaaaattg caagagacaa tggaccaggt cataatatta
     3661 catttcagag ttcacctccc acagttgagt ggcatataag cagacctggg cacatagaga
     3721 cttttgacct gctcacctta cacccaatag aaattgctcg acaactcact ttacttgaat
     3781 cagatctata ccgagctgta cagccatcag aattagttgg aagtgtgtgg acaaaagaag
     3841 acaaagaaat taactctcct aatcttctga aaatgattcg acataccacc aacctcactc
     3901 tgtggtttga gaaatgtatt gtagaaactg aaaatttaga agaaagagta gctgtggtga
     3961 gtcgaattat tgagattcta caagtctttc aagagttgaa caactttaat ggtgtccttg
     4021 aggttgtcag tgctatgaat tcatcacctg tttacagact agaccacaca tttgagcaaa
     4081 taccaagtcg ccagaagaaa attttagaag aagctcatga attgagtgaa gatcactata
     4141 agaaatattt ggcaaaactc aggtctatta atccaccatg tgtgcctttc tttggaattt
     4201 atctcactaa tatcttgaaa acagaagaag gcaaccctga ggtcctaaaa agacatggaa
     4261 aagagcttat aaactttagc aaaaggagga aagtagcaga aataacagga gagatccagc
     4321 agtaccaaaa tcagccttac tgtttacgag tagaatcaga tatcaaaagg ttctttgaaa
     4381 acttgaatcc gatgggaaat agcatggaga aggaatttac agattatctt ttcaacaaat
     4441 ccctagaaat agaaccacga aaccctaagc ctctcccaag atttccaaaa aaatatagct
     4501 atcccctaaa atctcctggt gttcgtccat caaacccaag accaggtacc taagatctta
     4561 aaaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     4621 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4681 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4741 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4801 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4861 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4921 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4981 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5041 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5101 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5161 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5221 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5401 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     5761 atcatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5821 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5881 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5941 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     6001 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6061 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6121 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6181 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6241 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6301 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6361 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6421 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6481 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6541 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6601 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6661 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6721 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6781 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6841 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6901 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6961 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     7021 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     7081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7321 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagt ttggagcgaa cgacctacac
     7561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7861 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     7921 cggggtcatt agttcatagc ccatatatgg agttccg
//
