LOCUS Exported 6103 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Recruitable YFP tagged RhoA GEF LARG (DH domain). Expresses in mammalian cells. ACCESSION . VERSION . KEYWORDS PR_GEF YFP (2XPDZ-YFP-LARG DH) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6103) AUTHORS Wagner E, Glotzer M TITLE Local RhoA activation induces cytokinetic furrows independent of spindle position and cell cycle stage. JOURNAL J Cell Biol. 2016 Jun 20;213(6):641-9. doi: 10.1083/jcb.201603025. Epub 2016 Jun 13. PUBMED 27298323 REFERENCE 2 (bases 1 to 6103) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6103 /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" misc_feature 1246..1298 /label=MCS /note="multiple cloning site" CDS 1300..2016 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1345..1366) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1606..1625) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1953..1974 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2891..3012 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2928..2947) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2982..3001 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3019..3474) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3156..3177) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3368..3387 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3501..3605 /gene="bla" /label=AmpR promoter promoter 3607..3964 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3630..3650) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3815..3950 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3877..3896 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3999..4793 /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(4053..4072) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4663..4682 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4981..5000) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5025..5072 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5401..5989 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5890..5909 /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 ccggactcag atctcgagct caagcttatc gataccgtcg acatggcaaa acaagagatt 661 cgagtgaggg ttgaaaagga cccagaactt ggatttagca tatcaggtgg tgtcgggggt 721 agaggaaacc cattcagacc tgatgatgat ggtatatttg taacaagggt acaacctgaa 781 ggaccagcat caaaattact gcagccaggt gataaaatta ttcaggctaa tggctacagt 841 tttataaata ttgaacatgg acaagcagtg tccttgctaa aaactttcca gaatacagtt 901 gaactcatca ttgtacgaga agttagtgct ggtggtagtg caggagggag tgctggcggg 961 gcaaaacaag agattcgagt gagggttgaa aaggacccag aacttggatt tagcatatca 1021 ggtggtgtcg ggggtagagg aaacccattc agacctgatg atgatggtat atttgtaaca 1081 agggtacaac ctgaaggacc agcatcaaaa ttactgcagc caggtgataa aattattcag 1141 gctaatggct acagttttat aaatattgaa catggacaag cagtgtcctt gctaaaaact 1201 ttccagaata cagttgaact catcattgta cgagaagttt ctagactcga gctcaagctt 1261 cgaattctgc aatcgacggt accgcgggcc cgggatccaa tggtgagcaa gggcgaggag 1321 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag 1381 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc 1441 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac 1501 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc 1561 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac 1621 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag 1681 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac 1741 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag 1801 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc 1861 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc 1921 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc 1981 gccgggatca ctctcggcat ggacgagctg tacaagctcg agctcaagct tagtgctggt 2041 ggtagtgcag gaggctcggc cggtggaagt gcaggaggga gtgctggcgg ggtcgacgac 2101 accccaccca actggcagca gcttgttagt cgagaagtgt tactgggact aaaaccttgt 2161 gaaatcaaaa gacaggaagt gattaatgaa ttgttctaca ctgaaagagc tcatgttcga 2221 acactgaagg ttcttgatca agtgttctat cagcgagtat ccagagaagg aattctgtca 2281 ccctcagagc tacggaaaat tttttcaaac ttggaagata ttcttcaact tcatattgga 2341 ttgaatgaac aaatgaaggc tgttcgaaag agaaatgaga cctctgttat cgatcagatt 2401 ggggaagatt tgctgacatg gttcagcgga ccaggagagg agaaattgaa acatgctgct 2461 gctacctttt gcagtaacca acctttcgcc ctggaaatga tcaaatctcg tcagaaaaag 2521 gattctcgat ttcagacttt tgtgcaagat gctgaaagta atccactgtg tcgtcgtctt 2581 caactgaagg atattattcc cactcaaatg caaaggctta ctaagtaccc acttctgttg 2641 gataatattg ccaaatacac agaatggcca acagaaaggg agaaggtgaa gaaagctgca 2701 gatcactgtc gtcagatctt aaattatgta aatcaggctg tcaaggaggc agaaaacaag 2761 cagcgctaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt 2821 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat 2881 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 2941 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 3001 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt 3061 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat 3121 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat 3181 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac 3241 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc 3301 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga 3361 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca 3421 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg 3481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 3541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 3601 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc 3661 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 3781 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 3841 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 3901 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 3961 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 4021 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 4081 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 4141 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat 4201 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 4261 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 4321 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 4381 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 4441 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 4501 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc 4561 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 4621 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 4681 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 4741 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 4801 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 4861 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 4921 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac 4981 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca 5041 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg 5101 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt 5161 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc 5221 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca 5281 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 5341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 5401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 5461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 5521 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 5581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 5641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 5701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 5761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 5821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 5881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 5941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6061 gttatcccct gattctgtgg ataaccgtat taccgccatg cat //