LOCUS Exported 5671 bp ds-DNA circular SYN 29-OCT-2021 DEFINITION Constitutively active, nuclear localized Rac1. ACCESSION . VERSION . KEYWORDS H2A-mTurquoise-RAC1-G12V-deltaCAAX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5671) AUTHORS Mahlandt EK, Arts JJG, van der Meer WJ, van der Linden FH, Tol S, van Buul JD, Gadella TWJ, Goedhart J TITLE Visualizing endogenous Rho activity with an improved localization-based, genetically encoded biosensor. JOURNAL J Cell Sci. 2021 Aug 6. pii: 271216. doi: 10.1242/jcs.258823. PUBMED 34357388 REFERENCE 2 (bases 1 to 5671) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 29, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5671 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 413..422 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 419..1135 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2010)" /label=mTurquoise /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(464..485) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(725..744) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1072..1093 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1830..1951 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1867..1886) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1921..1940 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1958..2413) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2095..2116) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2307..2326 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2440..2544 /gene="bla" /label=AmpR promoter promoter 2546..2903 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2569..2589) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2754..2889 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2816..2835 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2938..3732 /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(2992..3011) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3602..3621 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3920..3939) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3964..4011 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4340..4928 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4829..4848 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 5103..5406 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5407..5610 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5561..5581 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 aattctgatg tcgggacgcg gcaagcaggg aggcaaagct cgcgccaaag ccaagacccg 61 ctcttctcgt gccggtctcc agttccccgt gggccgagtg caccgactgc tccgcaaggg 121 caactatgct gagcgggtcg gggccggcgc gccggtgtac ctggcggcgg tgctggagta 181 cctgactgcc gagatcctgg agctggcggg caacgccgcc cgcgacaaca agaagacccg 241 cattatcccg cgccacttgc agctggccat ccgcaacgac gaggagctca acaagctgct 301 gggcaaagta accatcgctc agggtggtgt cctgcccaac atccaggctg tgctactgcc 361 caagaagacc gagagtcacc acaaggccaa aggcaaacgg gatccaccgg tcgccaccat 421 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 481 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 541 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 601 cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca 661 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt 721 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 781 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 841 gctggagtac aactacatca gcgacaacgt ctatatcacc gccgacaagc agaagaacgg 901 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga 961 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 1021 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 1081 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccat 1141 gcaggccatc aagtgtgtgg tggtgggaga cgtagctgta ggtaaaactt gcctactgat 1201 cagttacaca accaatgcat ttcctggaga atatatccct actgtctttg acaattattc 1261 tgccaatgtt atggtagatg gaaaaccggt gaatctgggc ttatgggata cagctggaca 1321 agaagattat gacagattac gccccctatc ctatccgcaa acagatgtgt tcttaatttg 1381 cttttccctt gtgagtcctg catcatttga aaatgtccgt gcaaagtggt atcctgaggt 1441 gcggcaccac tgtcccaaca ctcccatcat cctagtggga actaaacttg atcttaggga 1501 tgataaagac acgatcgaga aactgaagga gaagaagctg actcccatca cctatccgca 1561 gggtctagcc atggctaagg agattggtgc tgtaaaatac ctggagtgct cggcgctcac 1621 acagcgaggc ctcaagacag tgtttgacga agcgatccga gcagtcctct gcccgcctcc 1681 cgtgaagaag aggaagagaa aaagctaagc ggccgcgact ctagatcata atcagccata 1741 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga 1801 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca 1861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 1921 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg 1981 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc 2041 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt 2101 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc 2161 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg 2221 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga 2281 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg 2341 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg 2401 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 2461 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 2521 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc 2581 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 2641 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2701 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 2761 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 2821 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt 2881 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg 2941 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 3001 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 3061 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa 3121 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 3181 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 3241 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 3301 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 3361 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 3421 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc 3481 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 3541 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 3601 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 3661 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 3721 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc 3781 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt 3841 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc 3901 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 3961 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg 4021 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat 4081 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc 4141 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt 4201 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 4261 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 4321 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 4381 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 4441 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 4501 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 4561 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 4621 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 4681 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 4741 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 4801 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 4861 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 4921 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 4981 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc 5041 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 5101 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 5161 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 5221 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 5281 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 5341 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 5401 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg 5461 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 5521 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt 5581 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc 5641 taccggactc agatctcgag ctcaagcttc g //