LOCUS       sequence-269472-        6556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET biosensor. Fyn-tagged biosensor targeting to membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6556)
  AUTHORS   Ouyang M, Wan R, Qin Q, Peng Q, Wang P, Wu J, Allen M, Shi Y, Laub 
            S, Deng L, Lu S, Wang Y
  TITLE     Sensitive FRET Biosensor Reveals Fyn Kinase Regulation by 
            Submembrane Localization.
  JOURNAL   ACS Sens. 2019 Jan 25;4(1):76-86. doi: 10.1021/acssensors.8b00896. 
            Epub 2019 Jan 9.
  PUBMED    30588803
REFERENCE   2  (bases 1 to 6556)
  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/acssensors.8b00896"; journalName: "ACS Sens"; date: 
            "2019-01-25- 25"; volume: "4"; issue: "1"; pages: "76-86"
COMMENT     Sequence Label: Fyn-Fyn Biosensor
FEATURES             Location/Qualifiers
     source          1..6556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1787..1789,1790..1792,1793..2305)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     primer_bind     complement(1832..1853)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2093..2112)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2440..2461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2855..3565
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     LCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLEF
                     LTAAGITEGMNELYK"
     primer_bind     complement(3578..3597)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3643..3698
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3697..3716)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4059..4075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4059..4075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4072..4094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4083..4099
                     /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(4107..4113,4114..4131,4132..4137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4152..4173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4232..4427
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4278..4413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4433..4567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4470..4489)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4524..4543
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4635..4652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4806..5394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4886..4905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5565..6356,6357..6425))
                     /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     6188..6207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6426..6530)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     1741 tatgggctgt gtgcaatgta aggataaaga agcaacaaaa ctgacgatgg tgagcaaggg
     1801 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1861 ccacaggttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1921 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1981 gacctggggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2041 caagtccgcc atgcccgaag gctacgtcca ggagcgtacc atcttcttca aggacgacgg
     2101 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2161 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2221 ctacatcagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccca
     2281 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2341 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2401 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2461 gaccgccgcc cgcatgcatt ggtattttgg gaagatcact cgtcgggagt ccgagcggct
     2521 gctgctcaac cccgaaaacc cccggggaac cttcttggtc cgggagagcg agacgacaaa
     2581 aggtgcctat gccctctccg tttctgactt tgacaacgcc aaggggctca atgtgaagca
     2641 ctacaagatc cgcaagctgg acagcggcgg cttctacatc acctcacgca cacagttcag
     2701 cagcctgcag cagctggtgg cctactactc caaacatgct gatggcttgt gccaccgcct
     2761 gactaacgtc tgtgggtcta catctggatc tgggaagccg ggttctggtg agggttctga
     2821 gaagatcgag ggcacctacg gcgtcgtgga gctctctaaa ggtgaagaat tattcactgg
     2881 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
     2941 cggtgaaggt gaaggtgatg ctacgtacgg taaattgacc ttaaaattac tctgtactac
     3001 tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg gtgttcaatg
     3061 ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     3121 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     3181 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     3241 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     3301 ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa
     3361 cattgaagat ggtggtgttc aattagctga ccattatcaa caaaatactc caattggtga
     3421 tggtccagtc ttgttaccag acaaccatta cttatcctat caatctgcct tattcaaaga
     3481 tccaaacgaa aagagagacc acatggtctt gttagaattt ttgactgctg ctggtattac
     3541 cgagggtatg aatgaattgt acaaataagt cgacggatct ttttccctct gccaaaaatt
     3601 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3661 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     3721 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     3781 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     3841 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3901 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3961 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     4021 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     4081 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4141 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4201 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     4261 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4321 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4381 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     4441 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4501 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4561 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5701 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5761 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5821 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5881 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5941 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6001 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6061 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6121 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6181 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6241 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6301 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6361 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6421 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6541 cgaaaagtgc cacctg
//
