LOCUS       sequence-429116-        5561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mEGFP(A206K) fused to FKBP-FKBP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  AUTHORS   Hernandez-Candia CN, Brady BR, Harrison E, Tucker CL
  TITLE     A platform to induce and mature biomolecular condensates using 
            chemicals and light.
  JOURNAL   Nat Chem Biol. 2024 Jan 8. doi: 10.1038/s41589-023-01520-1.
  PUBMED    38191942
REFERENCE   2  (bases 1 to 5561)
  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"; journalName: "Nat Chem 
            Biol. 2024 Jan 8. doi: 10.1038/s41589-023-01520-1."
COMMENT     Sequence Label: B2387_mEGFP-FKBP2x
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(685..687,688..690,691..1401)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(730..751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(991..1010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1338..1359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1414..1734
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1840..2160
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    2349..2470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2386..2405)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2440..2459
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2477..2932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2614..2635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2826..2845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2959..3063
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3065..3422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3065..3256
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3088..3108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3273..3408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3457..4251
                     /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             3457..4251
                     /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(3511..3530)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4121..4140
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4439..4458)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4483..4530
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4859..5447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5348..5367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatgaacct ggtggacctc caaaagaagc tggaggagct ggagctggac gagcagcagg
      661 gaccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      841 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      901 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1081 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1141 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1201 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1261 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     1321 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1381 ggcatggacg agctgtacaa gagtgctggt ggtggagtgc aggtggaaac catctcccca
     1441 ggagacgggc gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg
     1501 cttgaagatg gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg
     1561 ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt
     1621 cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc
     1681 atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaagtgct
     1741 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtcctcga
     1801 gctagtgctg gtggtagtgc tggtggtagt gctggtggtg gagtgcaggt ggaaaccatc
     1861 tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt gcactacacc
     1921 gggatgcttg aagatggaaa gaaatttgat tcctcccggg acagaaacaa gccctttaag
     1981 tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt
     2041 gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac
     2101 ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct aaaactggaa
     2161 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatccag
     2221 cacagtggcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2281 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2341 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2401 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2461 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2521 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2581 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2641 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2701 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2761 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2821 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2881 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     2941 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3001 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3061 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3361 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3421 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3481 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3541 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3601 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3661 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3721 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3781 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3841 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3901 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3961 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4021 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4081 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4141 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4201 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4261 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4321 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4381 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4441 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4501 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4561 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4621 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4681 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4921 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4981 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5221 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5521 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
