LOCUS       sequence-426072-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-FKBP-FKBP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5552)
  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 5552)
  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: B2329_mCherry-FKBP2x
FEATURES             Location/Qualifiers
     source          1..5552
                     /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             685..1392
                     /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(830..848)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1098..1117
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1405..1725
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1831..2151
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    2340..2461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2377..2396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2431..2450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2468..2923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2605..2626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2817..2836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2950..3054
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3056..3413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3056..3247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3079..3099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3264..3399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3448..4242
                     /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             3448..4242
                     /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(3502..3521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4112..4131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4430..4449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4474..4521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4850..5438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5339..5358
                     /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 aggaggataa catggccatc
      721 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      781 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
      841 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
      901 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
      961 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1021 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1081 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1141 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1201 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1261 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     1321 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1381 gagctgtaca agagtgctgg tggtggagtg caggtggaaa ccatctcccc aggagacggg
     1441 cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat
     1501 ggaaagaaat ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag
     1561 caggaggtga tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc
     1621 aaactgacta tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca
     1681 ccacatgcca ctctcgtctt cgatgtggag cttctaaaac tggaaagtgc tggtggtagt
     1741 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggtcctcg agctagtgct
     1801 ggtggtagtg ctggtggtag tgctggtggt ggagtgcagg tggaaaccat ctccccagga
     1861 gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac cgggatgctt
     1921 gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa gtttatgcta
     1981 ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag tgtgggtcag
     2041 agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca cccaggcatc
     2101 atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga aaagcttggt
     2161 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca gcacagtggc
     2221 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2281 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2461 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2521 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2581 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2641 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2701 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2761 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2821 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2881 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     2941 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3001 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     3061 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     3421 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3481 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3541 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3601 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     3661 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3721 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3781 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3841 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3901 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3961 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4021 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4081 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4141 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4201 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4261 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4321 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4381 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     4441 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4501 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4561 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4621 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4681 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5521 attctgtgga taaccgtatt accgccatgc at
//
