LOCUS       sequence-175150-        6287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRB-tagged N-terminal Venus (YFP) fragment for BiFC; localizes to 
            plasma membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  AUTHORS   Hertel F, Mo GC, Duwe S, Dedecker P, Zhang J
  TITLE     RefSOFI for Mapping Nanoscale Organization of Protein-Protein 
            Interactions in Living Cells.
  JOURNAL   Cell Rep. 2016 Jan 12;14(2):390-400. doi: 
            10.1016/j.celrep.2015.12.036. Epub 2015 Dec 31.
  PUBMED    26748717
REFERENCE   2  (bases 1 to 6287)
  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: 
            "10.1016/j.celrep.2015.12.036"; journalName: "Cell Rep"; date: 
            "2016-01-12- 12"; volume: "14"; issue: "2"; pages: "390-400"
COMMENT     Sequence Label: pcDNA3-Lyn-FRB-Venus-173-N
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..318
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             join(355..357,358..360,361..870)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNI"
     primer_bind     complement(400..421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(661..680)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(934..952)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(935..952)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(972..989)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    978..1202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1248..1676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1335..1354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1545..1566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1685..1705)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1690..2019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1870..2005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1932..1951
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2086..2880
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2140..2159)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2750..2769
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3054..3175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3091..3110)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3145..3164
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3237..3259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3248..3264
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3272..3278,3279..3296,3297..3302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3317..3338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3455..3472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3626..4214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3706..3725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4385..5176,5177..5245))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5008..5027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5246..5350)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5425..5444)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5616..5995
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5996..6199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6150..6170
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6244..6263
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6244..6262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(6282..6287,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggctgca tcaagagcaa gcgcaaggac aaggatccca tcctctggca tgagatgtgg
       61 catgaaggcc tggaagaggc atctcgtttg tactttgggg aaaggaacgt gaaaggcatg
      121 tttgaggtgc tggagccctt gcatgctatg atggaacggg gcccccagac tctgaaggaa
      181 acatccttta atcaggccta tggtcgagat ttaatggagg cccaagagtg gtgcaggaag
      241 tacatgaaat cagggaatgt caaggacctc ctccaagcct gggacctcta ttatcatgtg
      301 ttccgacgaa tctcaaagac tagtggtacc ggtggaggta gtggaggtgg aagcatggtg
      361 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      421 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      481 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      541 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      601 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      661 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      721 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      781 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
      841 aaggccaact tcaagatccg ccacaacatc taagaattct gcagatatcc atcacactgg
      901 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
      961 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1021 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1081 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1141 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1201 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1441 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1561 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1681 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1741 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1861 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1921 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     1981 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2041 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2101 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2161 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2221 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2281 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2341 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2401 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2461 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2521 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2581 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     2641 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2701 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2761 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     2821 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     2881 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     2941 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3001 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3181 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3241 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3301 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3361 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5341 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5401 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5461 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     5521 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     5581 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     5641 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5701 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5761 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5821 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     5881 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     5941 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     6001 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6061 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6121 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     6181 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     6241 aattaatacg actcactata gggagaccca agcttgcggc cgccacc
//
