LOCUS       sequence-402910-        6724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP12-Talin(1974-2541)(K2443D)(V2444D)(K2445D)-mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6724)
  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 6724)
  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: FKBP12-talinC11-ABS3mut-mEmerald
FEATURES             Location/Qualifiers
     source          1..6724
                     /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             join(598..600,601..921)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      2662..2671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2668..2670,2671..2673,2674..3387)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2713..2734)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2974..2993)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3321..3342
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3509..3630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3600..3619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3637..4092)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3774..3795)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3986..4005
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4119..4223
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4225..4582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4225..4416
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4248..4268)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4433..4568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4617..5411
                     /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             4617..5411
                     /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(4671..4690)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5281..5300
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5599..5618)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5643..5690
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6019..6607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6508..6527
                     /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 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag
      661 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
      721 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
      781 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
      841 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
      901 gtggagcttc taaaactgga aactagtgat atcggtaccc aggcctgcat tacagcagcc
      961 agtgctgtgt ctggtatcat tgctgacctc gacaccacca tcatgttcgc tactgctggc
     1021 acacttaacc gtgagggtgc tgaaactttt gctgaccacc gggagggtat cttaaagaca
     1081 gcaaaggttc ttgtggagga caccaaggtc ctagtgcaga atgcagctgg gagccaggag
     1141 aagttggcac aagccgccca gtcctccgtg gccaccatta cccgcctcgc tgatgtggtc
     1201 aagctcggtg cagccagcct aggagccgaa gaccctgaaa ctcaggtggt gctgatcaat
     1261 gcagtaaagg acgtagccaa ggccctgggt gacctcatca gcgctacgaa ggctgcagcg
     1321 ggcaaagttg gggatgaccc tgcagtgtgg cagctcaaga actctgccaa ggtgatggtg
     1381 accaatgtga catcattgct caagacagtg aaggctgtgg aagatgaggc caccaaaggc
     1441 acacgggccc tagaggcaac cacagagcac atacgtcagg aactggcggt cttctgttcc
     1501 ccagagccac ctgccaagac ctctacccct gaagacttca tccgaatgac caagggtatt
     1561 actatggcaa cagccaaagc cgttgctgct ggcaattcct gtcgacagga agatgtcatt
     1621 gccacagcca atctgagccg acgggctatt gcggacatgc ttcgggcttg caaggaagca
     1681 gctttccacc cagaagtggc gcctgatgta cggctccgag ccctgcacta tggccgggag
     1741 tgtgccaatg gttacctgga actgctggac cacgtgctgc tgacccttca gaagccaaac
     1801 ccagacctga agcagcagct aactggacac tcaaagcggg ttgctggctc tgtgactgag
     1861 ctcatccagg ctgctgaagc catgaaagga acagagtggg tggacccaga ggaccctact
     1921 gtcattgctg agaatgagct cttgggagcc gcagccgcca tcgaggctgc agccaagaag
     1981 ctggagcagc tgaagcccag ggccaaaccc aaggaggcgg atgagtcctt gaactttgag
     2041 gaacaaatcc tagaagctgc caagtccatc gctgcagcca ccagtgcact ggtaaaggct
     2101 gcgtcagcag cccagaggga gctggtggct caaggaaagg tgggcgccat tccagccaat
     2161 gcactggatg atgggcagtg gtcgcagggc ctcatttccg ctgcccgtat ggtggctgcg
     2221 gccaccaaca atctgtgtga agcagctaac gcagctgtcc agggccatgc tagccaggag
     2281 aaactcattt cctcagccaa gcaagtagct gcctccacag ctcagctcct ggtagcttgc
     2341 gatgacgatg cagatcagga ctctgaggca atgaaacggc tccaggctgc tggcaatgca
     2401 gtgaagaggg cctcagataa cctggtaaag gcggcccaga aagctgcagc cttcgaagac
     2461 caggagaatg agacggtggt ggtgaaggag aaaatggttg ggggcattgc ccagattatc
     2521 gcagcacagg aagagatgct tcggaaggaa cgagagctgg aagaggctcg gaaaaagctc
     2581 gcccagatcc ggcagcagca gtacaagttc ttgccttcag agcttcgaga cgagcacggc
     2641 tccgcgtccc gatcaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2701 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2761 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2821 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tgacctacgg cgtgcagtgc
     2881 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2941 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3001 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3061 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaaggtc
     3121 tatatcaccg ccgacaagca gaagaacggc atcaaggtga acttcaagac ccgccacaac
     3181 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3241 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     3301 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3361 ctcggcatgg acgagctgta caagtaagcg gccgcgactc tagatcataa tcagccatac
     3421 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3481 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3541 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3601 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3661 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3721 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3781 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3841 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3901 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3961 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4021 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4081 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4141 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4201 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4261 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4321 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4381 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4441 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4501 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4561 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4621 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4681 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4741 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4801 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4861 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4921 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4981 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5041 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5101 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5161 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5221 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5281 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5341 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5401 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5461 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5521 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5581 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5641 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5701 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5761 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5821 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5881 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5941 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6721 ttag
//
