LOCUS       sequence-266126-        7499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively-active (CA) mutant of OsRac1; Acts as a positive 
            control for small GTPases activation assays..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7499)
  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 7499)
  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 CA
FEATURES             Location/Qualifiers
     source          1..7499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1992
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    2024..2048
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2096..2098,2099..2101,2102..2614)
                     /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(3617..3619,3620..3622,3623..4135)
                     /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"
     protein_bind    complement(4543..4567)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      4599..4851
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4868..4884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4868..4884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4881..4903)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4892..4908
                     /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(4916..4922,4923..4940,4941..4946))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4961..4982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5099..5116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5270..5858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5350..5369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6029..6820,6821..6889))
                     /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     6652..6671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6890..6994)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7062..7080
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7118..7140)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7240..7259
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7453..7475
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7467..7484
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7468..7484
                     /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 actagagtta tcaacaagtt tgtacaaaaa
     2041 agcaggctct ttaaaggaac caattcagtc gactggatcc ggtaccgaat tcggcatggt
     2101 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2161 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2221 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2281 gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     2341 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2401 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2461 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2521 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     2581 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     2641 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2701 gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2761 ggagttcgtg accgccgccc tcgagaaaga gaaagagcgg ccagagattt ctctcccttc
     2821 agattttgaa cacacaattc atgtcggttt tgatgctgtc acaggggagt ttacgggaat
     2881 gccagagcag tgggcccgct tgcttcagac atcaaatatc actaagtcgg agcagaagaa
     2941 aaacccgcag gctgttctgg atgtgttgga gttttacaac tcgaagaaga catccaacag
     3001 ccagaaatac atgagcttta cagataagtc agcttccgga ggtggaaccg gtggtggagg
     3061 taccatgagc tcggcggcgg cggcgacgag gttcatcaag tgcgtcaccg tgggggacgt
     3121 cgcggtgggg aagacgtgca tgctcatctg ctacacctgc aacaagttcc ccaccgatta
     3181 catccccacc gtgttcgaca acttcagcgc caatgtctcc gtggacggga gcgtcgtcaa
     3241 cctcggcctc tgggacactg caggtcagga ggattacagc aggttgaggc ctctgagcta
     3301 caggggagcc gatgtgttca tcctgtcctt ctccctgata agcagggcga gctatgagaa
     3361 tgttcagaag aagtggatgc cagagcttcg ccggtttgcg cctggtgttc ctgtagttct
     3421 tgttggaacc aagttggatc tccgtgaaga tagggcctat cttgctgatc atccagcttc
     3481 ttccataata acaacggagc agggagaaga actgaggaag ctaataggag cggtcgccta
     3541 catcgaatgc agctccaaga cacagagaaa cattaaagct gttttcgaca ctgccatcaa
     3601 agtggtgcgc ggccgcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3661 cctggtcgag ctggacggcg acgtaaacgg ccacaggttc agcgtgtccg gcgagggcga
     3721 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3781 cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct tcagccgcta
     3841 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3901 ggagcgtacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3961 cgagggcgac accctggtga accgcatcga gctgaagggc atcggcttca aggaggacgg
     4021 caacatcctg gggcacaagc tagagtacaa ctacatcagc cacaacgtct atatcaccgc
     4081 cgacaagcag aagaacggca tcaaggccca cttcaagatc cgccacaaca tcgaggacgg
     4141 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4201 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4261 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4321 cgagctgtct agaccaagac ataaggatgt aaccagaaag aaactccaat caagctccaa
     4381 tcggccagta aggcggtact tttgcggaag cgcttgtttc gcgtagggat cctctagacc
     4441 aagacataag gatgtaacca gaaagaaact ccaatcaagc tccaatcggc cagtaaggcg
     4501 gtacttttgc ggaagcgctt gtttcgcgta gggatcctct agacccagct ttcttgtaca
     4561 aagtggttga taacagcgct tagagctcga atttccccga tcgttcaaac atttggcaat
     4621 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     4681 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     4741 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     4801 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattcg
     4861 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4921 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4981 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5041 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5101 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5161 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5221 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5281 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5341 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5401 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5461 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5521 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5581 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5641 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5701 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5761 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5821 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5881 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5941 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6001 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6061 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6121 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6181 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6241 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6301 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6361 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6421 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6481 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6541 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6601 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6661 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6721 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6781 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6841 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6901 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6961 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7021 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7081 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7141 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7201 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7261 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7321 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     7381 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     7441 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgcatgc
//
