LOCUS       sequence-426081-        6241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses miRFP-FKBP-FKBP-FKBP-FKBP-FKBP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6241)
  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 6241)
  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: B2401_miRFP-FKBP5x
FEATURES             Location/Qualifiers
     source          1..6241
                     /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      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1065..1385
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1491..1811
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1842..2162
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2205..2525
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2568..2888
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3029..3150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3066..3085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3120..3139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3157..3612)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3294..3315)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3506..3525
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3639..3743
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3745..4102
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3745..3936
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3768..3788)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3953..4088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4137..4931
                     /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             4137..4931
                     /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(4191..4210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4801..4820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5119..5138)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5163..5210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5539..6127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6028..6047
                     /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 ccatggcaaa cctggacaag atgctgaata ccacagtaac agaggtgcgg cagttcctgc
      661 aggtggacag agtgtgcgtg ttccagtttg aggaggatta tagcggagtg gtggtggtgg
      721 aggccgtgga cgataggtgg atctccatcc tgaagaccca ggtgcgggat agatacttca
      781 tggagacaag gggcgaggag tattctcacg gccgctacca ggccatcgcc gacatctaca
      841 ccgcaaacct gacagagtgc tacagggatc tgctgacaca gtttcaggtg agagcaatcc
      901 tggccgtgcc catcctgcag ggcaagaagc tgtggggcct gttggtggca caccagctgg
      961 cggcccctag acagtggcag acctgggaga tcgactttct gaagcagcag gccgtggtgg
     1021 tgggcatcgc catccagcag agcgtgtaca agagtgctgg tggtggagtg caggtggaaa
     1081 ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact
     1141 acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga aacaagccct
     1201 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     1261 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     1321 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     1381 tggaaagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     1441 gtggtcctcg agctagtgct ggtggtagtg ctggtggtag tgctggtggt ggagtgcagg
     1501 tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg
     1561 tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca
     1621 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     1681 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     1741 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     1801 taaaactgga aaagcttagt gctggtggta gtgctggtgg tggagtgcag gtggaaacca
     1861 tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca
     1921 ccgggatgct tgaagatgga aagaaatttg attcctcccg ggacagaaac aagcccttta
     1981 agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga
     2041 gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc
     2101 acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg
     2161 aagaattcag tgctggtggt agtgctggtg gtagtgctgg tggtggagtg caggtggaaa
     2221 ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact
     2281 acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga aacaagccct
     2341 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     2401 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     2461 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     2521 tggaagtcga cagtgctggt ggtagtgctg gtggtagtgc tggtggtgga gtgcaggtgg
     2581 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
     2641 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
     2701 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
     2761 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
     2821 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
     2881 aactggaacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     2941 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3001 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3061 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3121 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3181 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3241 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3301 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3361 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3421 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3481 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3541 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3601 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3661 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3721 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3781 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3901 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3961 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4021 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4081 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4141 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4201 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4261 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4321 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4381 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4441 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4501 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4561 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4621 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4681 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4741 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4801 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4861 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4921 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4981 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5041 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5101 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5161 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5221 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5281 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5341 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5401 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5461 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5521 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5581 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5641 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5701 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5761 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5821 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5881 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5941 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6001 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6061 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6121 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6181 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6241 t
//
