LOCUS       sequence-266091-        7285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosensor for monitoring activation for rice small GTPase OsRac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7285)
  AUTHORS   Wong HL, Akamatsu A, Wang Q, Higuchi M, Matsuda T, Okuda J, Kosami 
            KI, Inada N, Kawasaki T, Kaneko-Kawano T, Nagawa S, Tan L, Kawano Y,
            Shimamoto K
  TITLE     In vivo monitoring of plant small GTPase activation using a Forster 
            resonance energy transfer biosensor.
  JOURNAL   Plant Methods. 2018 Jul 7;14:56. doi: 10.1186/s13007-018-0325-4. 
            eCollection 2018.
  PUBMED    30002723
REFERENCE   2  (bases 1 to 7285)
  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: "Plant 
            Methods."; date: "2018-07-7"; volume: "14:56. doi"; pages: " 
            10.1186/s13007-018-0325-4. eCollection 2018"
COMMENT     Sequence Label: Raichu-OsRac1 WT
FEATURES             Location/Qualifiers
     source          1..7285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1992
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             join(2020..2022,2023..2025,2026..2538)
                     /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
                     ANFKIRHNIE"
     CDS             join(3541..3543,3544..3546,3547..4059)
                     /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
                     GNYKTRAEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     terminator      4385..4637
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4654..4670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4654..4670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4667..4689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4678..4694
                     /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(4702..4708,4709..4726,4727..4732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4747..4768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4885..4902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5056..5644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5136..5155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5815..6606,6607..6675))
                     /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     6438..6457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6676..6780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6848..6866
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6904..6926)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7026..7045
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7239..7261
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7253..7270
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7254..7270
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctgcagtgca gcgtgacccg gtcgtgcccc tctctagaga taatgagcat tgcatgtcta
       61 agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc agtttatcta
      121 tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt actacaataa
      181 tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa ggacaattga
      241 gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt tctccttttt
      301 ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca tccatttagg
      361 gtttagggtt aatggttttt atagactaat ttttttagta catctatttt attctatttt
      421 agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat aatttagata
      481 taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag aaattaaaaa
      541 aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa acgccgtcga
      601 cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa gcgaagcaga
      661 cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct ccaccgttgg
      721 acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt gagccggcac
      781 ggcaggcggc ctcctcctcc tctcacggca ccggcagcta cgggggattc ctttcccacc
      841 gctccttcgc tttcccttcc tcgcccgccg taataaatag acaccccctc cacaccctct
      901 ttccccaacc tcgtgttgtt cggagcgcac acacacacaa ccagatctcc cccaaatcca
      961 cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc cccccccccc ctctctacct
     1021 tctctagatc ggcgttccgg tccatggtta gggcccggta gttctacttc tgttcatgtt
     1081 tgtgttagat ccgtgtttgt gttagatccg tgctgctagc gttcgtacac ggatgcgacc
     1141 tgtacgtcag acacgttctg attgctaact tgccagtgtt tctctttggg gaatcctggg
     1201 atggctctag ccgttccgca gacgggatcg atttcatgat tttttttgtt tcgttgcata
     1261 gggtttggtt tgcccttttc ctttatttca atatatgccg tgcacttgtt tgtcgggtca
     1321 tcttttcatg cttttttttg tcttggttgt gatgatgtgg tctggttggg cggtcgttct
     1381 agatcggagt agaattctgt ttcaaactac ctggtggatt tattaatttt ggatctgtat
     1441 gtgtgtgcca tacatattca tagttacgaa ttgaagatga tggatggaaa tatcgatcta
     1501 ggataggtat acatgttgat gcgggtttta ctgatgcata tacagagatg ctttttgttc
     1561 gcttggttgt gatgatgtgg tgtggttggg cggtcgttca ttcgttctag atcggagtag
     1621 aatactgttt caaactacct ggtgtattta ttaattttgg aactgtatgt gtgtgtcata
     1681 catcttcata gttacgagtt taagatggat ggaaatatcg atctaggata ggtatacatg
     1741 ttgatgtggg ttttactgat gcatatacat gatggcatat gcagcatcta ttcatatgct
     1801 ctaaccttga gtacctatct attataataa acaagtatgt tttataatta ttttgatctt
     1861 gatatacttg gatgatggca tatgcagcag ctatatgtgg atttttttag ccctgccttc
     1921 atacgctatt tatttgcttg gtactgtttc ttttgtcgat gctcaccctg ttgtttggtg
     1981 ttacttctgc aggtcgactc tagaggatcc gaattcggca tggtgagcaa gggcgaggag
     2041 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2101 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     2161 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     2221 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2281 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2341 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2401 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2461 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     2521 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     2581 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2641 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2701 gccctcgaga aagagaaaga gcggccagag atttctctcc cttcagattt tgaacacaca
     2761 attcatgtcg gttttgatgc tgtcacaggg gagtttacgg gaatgccaga gcagtgggcc
     2821 cgcttgcttc agacatcaaa tatcactaag tcggagcaga agaaaaaccc gcaggctgtt
     2881 ctggatgtgt tggagtttta caactcgaag aagacatcca acagccagaa atacatgagc
     2941 tttacagata agtcagcttc cggaggtgga accggtggtg gaggtaccat gagctcggcg
     3001 gcggcggcga cgaggttcat caagtgcgtc accgtggggg acggcgcggt ggggaagacg
     3061 tgcatgctca tctgctacac ctgcaacaag ttccccaccg attacatccc caccgtgttc
     3121 gacaacttca gcgccaatgt ctccgtggac gggagcgtcg tcaacctcgg cctctgggac
     3181 actgcaggtc aggaggatta cagcaggttg aggcctctga gctacagggg agccgatgtg
     3241 ttcatcctgt ccttctccct gataagcagg gcgagctatg agaatgttca gaagaagtgg
     3301 atgccagagc ttcgccggtt tgcgcctggt gttcctgtag ttcttgttgg aaccaagttg
     3361 gatctccgtg aagatagggc ctatcttgct gatcatccag cttcttccat aataacaacg
     3421 gagcagggag aagaactgag gaagctaata ggagcggtcg cctacatcga atgcagctcc
     3481 aagacacaga gaaacattaa agctgttttc gacactgcca tcaaagtggt gcgcggccgc
     3541 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3601 ggcgacgtaa acggccacag gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3661 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3721 ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3781 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg taccatcttc
     3841 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3901 gtgaaccgca tcgagctgaa gggcatcggc ttcaaggagg acggcaacat cctggggcac
     3961 aagctagagt acaactacat cagccacaac gtctatatca ccgccgacaa gcagaagaac
     4021 ggcatcaagg cccacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     4081 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4141 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4201 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtctagacca
     4261 agacataagg atgtaaccag aaagaaactc caatcaagct ccaatcggcc agtaaggcgg
     4321 tacttttgcg gaagcgcttg tttcgcgtag ggatccacta gtggtaccga gctcgaattt
     4381 ccccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     4441 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     4501 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     4561 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     4621 atctatgtta ctagatcggg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     4681 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5461 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6661 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6781 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6841 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6901 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6961 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7021 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     7081 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     7141 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7201 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     7261 gacggccagt gccaagcttg catgc
//
