LOCUS Exported 5271 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Rac1 FLIM donor for dynamic imaging of GTPase activity. ACCESSION . VERSION . KEYWORDS mEGFP-Rac1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5271) AUTHORS Hedrick NG, Harward SC, Hall CE, Murakoshi H, McNamara JO, Yasuda R TITLE Rho GTPase complementation underlies BDNF-dependent homo- and heterosynaptic plasticity. JOURNAL Nature. 2016 Sep 28;538(7623):104-108. doi: 10.1038/nature19784. PUBMED 27680697 REFERENCE 2 (bases 1 to 5271) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5271 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" CDS 1351..1929 /codon_start=1 /product="human Rac family small GTPase 1" /label=Rac1 /translation="MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVM VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR GLKTVFDEAIRAVLCPPPVKKRKRKCLLL" polyA_signal 2059..2180 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2096..2115) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2150..2169 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2187..2642) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2324..2345) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2536..2555 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2669..2773 /gene="bla" /label=AmpR promoter promoter 2775..3132 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2798..2818) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2983..3118 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3045..3064 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3167..3961 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3221..3240) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3831..3850 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4149..4168) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4193..4240 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4569..5157 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5058..5077 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgaggg atgcaggcca tcaagtgtgt ggtggtggga 1381 gacggagctg taggtaaaac ttgcctactg atcagttaca caaccaatgc atttcctgga 1441 gaatatatcc ctactgtctt tgacaattat tctgccaatg ttatggtaga tggaaaaccg 1501 gtgaatctgg gcttatggga tacagctgga caagaagatt atgacagatt acgcccccta 1561 tcctatccgc aaacagatgt gttcttaatt tgcttttccc ttgtgagtcc tgcatcattt 1621 gaaaatgtcc gtgcaaagtg gtatcctgag gtgcggcacc actgtcccaa cactcccatc 1681 atcctagtgg gaactaaact tgatcttagg gatgataaag acacgatcga gaaactgaag 1741 gagaagaagc tgactcccat cacctatccg cagggtctag ccatggctaa ggagattggt 1801 gctgtaaaat acctggagtg ctcggcgctc acacagcgag gcctcaagac agtgtttgac 1861 gaagcgatcc gagcagtcct ctgcccgcct cccgtgaaga agaggaagag aaaatgcctg 1921 ctgttgtaag gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta 1981 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 2041 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 2101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 2161 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc 2221 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 2281 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 2341 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 2401 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 2461 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 2521 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 2581 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 2641 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 2701 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 2761 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 2821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2881 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2941 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3001 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3061 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3121 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 3181 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 3241 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 3301 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 3361 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 3421 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 3481 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 3541 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 3601 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 3661 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 3721 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 3781 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 3841 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 3901 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 3961 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 4021 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 4081 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 4141 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 4201 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 4261 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 4321 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 4381 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 4441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 4501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 4561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 4621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 4681 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 4741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 4801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 4861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 4921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 4981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 5161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 5221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t //