LOCUS Exported 7285 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Dominant-negative (DN) mutant of OsRac1; Acts as a negative control for small GTPases activation assays. ACCESSION . VERSION . KEYWORDS Raichu-OsRac1 DN 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7285 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..1992 /label=Ubi promoter /note="maize polyubiquitin gene promoter" CDS 2020..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" 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(4702..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(5815..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 ggggaagaac 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 aaggaaactc 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 //