LOCUS       sequence-402906-        7681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mApple-Talin(1-911)-FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Wang Y, Barnett SFH, Le S, Guo Z, Zhong X, Kanchanawong P, Yan J
  TITLE     Label-free Single-Molecule Quantification of Rapamycin-induced 
            FKBP-FRB Dimerization for Direct Control of Cellular 
            Mechanotransduction.
  JOURNAL   Nano Lett. 2019 Oct 9;19(10):7514-7525. doi: 
            10.1021/acs.nanolett.9b03364. Epub 2019 Sep 6.
  PUBMED    31466449
REFERENCE   2  (bases 1 to 7681)
  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/acs.nanolett.9b03364"; journalName: "Nano Lett"; date: 
            "2019-10-9- 9"; volume: "19"; issue: "10"; pages: "7514-7525"
COMMENT     Sequence Label: mApple-TalinN3-FRB(wt)
FEATURES             Location/Qualifiers
     source          1..7681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      592..601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             598..1305
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1011..1030
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4063..4341
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     polyA_signal    4466..4587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4503..4522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4557..4576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4594..5049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4731..4752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4943..4962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5076..5180
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5182..5539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5182..5373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5205..5225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5390..5525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5574..6368
                     /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             5574..6368
                     /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(5628..5647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6238..6257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6556..6575)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6600..6647
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6976..7564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7465..7484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agccaccatg
      601 gtgagcaagg gcgaggagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg
      661 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      721 ccctacgagg cctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc
      781 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggtctacat taagcaccca
      841 gccgacatcc ccgactactt caagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg
      901 atgaacttcg aggacggcgg cattattcac gttaaccagg actcctccct gcaggacggc
      961 gtgttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1021 cagaagaaga ccatgggctg ggaggcctcc gaggagcgga tgtaccccga ggacggcgcc
     1081 ctgaagagcg agatcaagaa gaggctgaag ctgaaggacg gcggccacta cgccgccgag
     1141 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcgac
     1201 atcaagttgg acatcgtgtc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     1261 gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggtcga cctcgagatg
     1321 gttgcgcttt cgctgaagat tagcattggg aatgtggtga agacgatgca atttgagcca
     1381 tctaccatgg tgtacgacgc ctgccgcatg attcgtgagc ggatcccaga ggccctggct
     1441 ggccctccca acgactttgg gctctttctg tcagatgatg accccaaaaa aggcatctgg
     1501 ctagaggctg ggaaagcttt ggactactac atgctccgaa atggggacac catggagtac
     1561 agaaaaaagc agagacccct gaagatacgg atgttagacg gaacggtgaa gactatcatg
     1621 gtggatgact ccaagaccgt cacagacatg ctcatgacca tttgtgcccg aattggtatc
     1681 accaaccatg atgagtattc actggttcga gagctgatgg aagaaaagaa agatgagggg
     1741 acagggaccc tgagaaaaga caagacccta ctgcgagatg aaaagaagat ggagaagcta
     1801 aagcagaagt tgcacacaga cgatgagttg aactggctgg accatgggcg gacactgagg
     1861 gaacagggag tggaagagca tgagacgttg ctgctgcgga gaaagttctt ctactcagac
     1921 cagaatgtgg attcccgaga ccctgtacag ctgaaccttc tctatgtgca ggcacgagat
     1981 gacatcctga atggctccca tcctgtctcc tttgacaagg cctgtgaatt tgcaggcttc
     2041 cagtgccaga tccagtttgg acctcacaat gaacagaagc acaaggctgg cttccttgac
     2101 ctgaaggact tcctgcccaa ggagtatgtg aagcagaagg gagagcgtaa gatctttcag
     2161 gcacacaaga attgtgggca gatgagtgaa attgaggcca aggtacgcta cgtgaagctg
     2221 gcccgttccc tcaagactta cggtgtctcc ttcttcctag tcaaggaaaa gatgaagggg
     2281 aagaataaac tggtgcccag gctgttgggc atcactaagg agtgtgtgat gcgtgtggat
     2341 gagaagacca aggaggtgat tcaggaatgg agcctcacca acatcaaacg ctgggctgcc
     2401 tctcccaaga gctttactct ggactttgga gactatcagg atggctacta ctcagtacag
     2461 acgacagaag gcgagcagat cgcacagctc attgctggct acatagatat catccttaag
     2521 aagaaaaaaa gcaaggacca ttttgggctg gagggagatg aagagtctac tatgttggag
     2581 gactcagtgt ctcccaaaaa gtcaacagtc cttcagcagc agtacaaccg agtggggaaa
     2641 gtggagcacg gctctgtggc tctgccagcc atcatgcgct ctggagcctc tggtcctgag
     2701 aatttccagg tggggagcat gccacctgcc cagcagcaga tcaccagtgg ccagatgcac
     2761 cgaggacaca tgccacctct gacttcagcc cagcaggcgc tcactggaac cattaactcc
     2821 agcatgcagg ctgtgcaggc tgcccaggcc actctggatg actttgagac tctaccccct
     2881 cttggccagg atgctgcctc caaagcttgg cgtaagaaca agatggatga atcaaagcat
     2941 gagatccact cccaggtaga tgccatcaca gctggcactg cctctgtggt aaacctgaca
     3001 gcaggagatc ctgcagagac agactataca gcagtgggct gtgcagtcac caccatctct
     3061 tccaacctga cggagatgtc ccgtggggtg aagctattgg ctgccctgct ggaggatgaa
     3121 ggcggcaatg gccggcccct tctgcaagca gcaaagggcc ttgcaggggc tgtgtcggaa
     3181 ctgcttcgca gtgctcagcc cgccagcgct gagcctcgtc agaacctgct gcaggcagcc
     3241 gggaacgtgg gccaggccag tggggagctg ttgcagcaaa ttggggaaag tgacactgac
     3301 ccccacttcc aggatgttct aatgcagcta gcaaacgcag tggcaagtgc tgcagccgcc
     3361 ctggtcctca aggccaagag tgtggcccag cgcacagagg attccgggct tcagacccaa
     3421 gtgattgctg cagctacgca atgtgctctg tccacttccc agctggtggc ctgtaccaag
     3481 gtggtagcac ctacaatcag ctcacctgtg tgccaagaac agctagtcga ggctgggcga
     3541 ctggtggcca aagctgtgga gggctgtgtg tctgcctcgc aagcagctac agaagatgga
     3601 cagctgcttc gaggggtagg agcagcagcc acggctgtca cccaggccct caatgagctg
     3661 ctgcagcacg tgaaggccca cgccaccgga gctgggcctg ccggtcggta tgaccaagcc
     3721 actgatacca tcctcaccgt cactgaaaac atcttcagct ccatgggtga tgctggggag
     3781 atggtgcgcc aggcccgcat cctggcccaa gccacctcag acctggtcaa tgctatcaag
     3841 gccgatgctg agggggagag tgatctggag aactctagaa agctcctgag tgctgccaag
     3901 atcctcgctg atgccaccgc caagatggtg gaggcggcca agggagcagc cgcccaccct
     3961 gacagtgagg aacagcagca gcgactgcgt gaagcagctg aggggcttcg catggccacc
     4021 aatgcagctg cgcagaacgc catcaagaag gaattcaccg gtatcctctg gcatgagatg
     4081 tggcatgaag gcctggaaga ggcatctcgt ttgtactttg gggaaaggaa cgtgaaaggc
     4141 atgtttgagg tgctggagcc cttgcatgct atgatggaac ggggccccca gactctgaag
     4201 gaaacatcct ttaatcaggc ctatggtcga gatttaatgg aggcccaaga gtggtgcagg
     4261 aagtacatga aatcagggaa tgtcaaggac ctcactcaag cctgggacct ctattatcat
     4321 gtgttccgac gaatctcaaa gtaagcggcc gcgactctag atcataatca gccataccac
     4381 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4441 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4501 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4561 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     4621 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4681 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4741 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4801 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4861 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4921 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4981 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5041 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5101 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5161 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5221 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5281 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5341 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5401 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5461 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5521 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5581 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5641 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5701 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5761 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5821 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5881 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5941 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6001 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6061 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     6121 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6181 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6241 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6301 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6361 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6421 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6481 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6541 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6601 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6661 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6721 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6781 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6841 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6901 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6961 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7021 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7081 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7141 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7201 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7261 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7321 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7381 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7441 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7501 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7561 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7621 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     7681 g
//
