LOCUS Exported 7588 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION RhoGEF. ACCESSION . VERSION . KEYWORDS mCherry-FGD1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7588) AUTHORS Reinhard NR, Van Der Niet S, Chertkova A, Postma M, Hordijk PL, Gadella TWJ Jr, Goedhart J TITLE Identification of guanine nucleotide exchange factors that increase Cdc42 activity in primary human endothelial cells. JOURNAL Small GTPases. 2019 Aug 30:1-15. doi: 10.1080/21541248.2019.1658509. PUBMED 31469028 REFERENCE 2 (bases 1 to 7588) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7588 /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..1320 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(758..776) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1026..1045 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 4376..4497 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4413..4432) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4467..4486 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4504..4959) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4641..4662) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4853..4872 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4986..5090 /gene="bla" /label=AmpR promoter promoter 5092..5449 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5115..5135) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5300..5435 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5362..5381 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5484..6278 /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(5538..5557) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6148..6167 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6466..6485) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6510..6557 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6886..7474 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7375..7394 /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 gaggataaca tggccatcat caaggagttc 661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt 781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc 841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc 901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc 1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac 1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc 1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc 1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg 1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag 1321 tccggactca gatctcatgg ccaccgagtc ccggggggcc ctgggccttc ggatcccgaa 1381 cgctcagcgg caaatacccc cggtgccgct ccgctggcct gtgccgactc ggaccctgga 1441 gccttggaac ccgggctgcc ggtgagcagg ggctcaggca cggctcttgg aggccctttg 1501 gatcctcagt ttgttggacc ctcggatgcc agcctgggag cgcctccaag ctcccgggtc 1561 ttaccctgcg gccctagtcc tcagcaccac cgggccctgc gcttctccta ccacctggag 1621 ggctcgcagc ctcggcctgg actgcaccag ggaaaccgga tcctggttaa aagtttgtcc 1681 cttgaccctg gccagagcct tgagcctcat ccagaaggtc cccaacggct tcgctcagat 1741 ccaggccctc ccactgaaat ccctggccca cgtccttcac cactgaagcg ggcaccgggc 1801 ccaaagccac aggttccccc aaagcccagc tacctgcaga tgccccgggt gcttcctcct 1861 cctgagccca ttcctccacc accatcacgc cctctgcctg cagaccctag agtggccaag 1921 ggactcgtcc ccagagctga ggccagcact agttctgcag cagtatcatc attgattgag 1981 aagtttgaaa gagagcctgt gattgttgcc tcggataggc cagcccctgg cccctgcccg 2041 gttcccccag agccagccat gttgccacag ccacccccgc agccaacagg gtcccagctc 2101 cctgagggtg aagcttctcg ctgcctgttc ctgctggctc ctgggccccg ggatggtgag 2161 aaggtgccta accgggacag cggcattgac agcatcagct cgccatccaa cagtgaagag 2221 acctgcttcg tcagtgatga tgggcccccc atccacagcc tctgtcctgg gccccccgcc 2281 ctggctagta tgccagtagc tttggccgac ccccaccggc ctggctccca ggaggttgac 2341 agtgacctgg aagaggaaga ggaagaagaa gaggaggaaa aggaaagaga aattccagtg 2401 cccccgatgg agaggcagga gtctgtggag ctgactgtgc agcagaaggt gttccatatt 2461 gccaatgagc tcctgcaaac tgagaaggcc tatgtttcca ggctccatct cctggatcag 2521 gtattctgtg cccggctgct ggaagaagct cggaaccgca gttccttccc cgcagatgtt 2581 gtccacggca tcttctctaa catctgctcc atctattgct tccaccagca gttcctgctg 2641 cctgagctag agaagcgcat ggaggaatgg gaccgctacc cacgcattgg tgacattctt 2701 cagaagttgg cacccttcct caagatgtat ggcgagtatg tgaaaaactt cgaccgggcg 2761 gtagagctgg tcaacacatg gacagagcgg tctacccagt ttaaagtcat catccatgaa 2821 gtacagaagg aggaagcctg tggcaacctg accttgcagc accacatgct ggagcccgtg 2881 cagcgtatcc cccgctatga acttcttctc aaggactatc tgttaaagct gcctcatggc 2941 tccccggaca gcaaggatgc ccagaagtct ttggagctga tagccacagc agcagaacac 3001 tctaatgctg ccatccggaa aatggaacga atgcacaagc tgctgaaggt gtatgagctg 3061 ctagggggtg aggaggacat tgtcagtccc accaaagagc tcataaagga aggccatatc 3121 cttaagctgt cagcaaagaa tgggaccact caagatcgat acctcatatt attcaacgac 3181 cgcctccttt actgcgtgcc caggctgcgg ctccttggtc agaagtttag cgtgcgggca 3241 cgcattgatg tagatggcat ggagctaaag gagagctcca acctcaacat gcctcggacc 3301 ttcctggtgt caggaaagca gcgctctctc gaactgcagg ccaggactga ggaggagaag 3361 aaagactggg tccaggccat caattccacc ctcctgaagc atgaacagac cctggagact 3421 ttcaagctgt tgaactcaac aaacagggat gatgaagata ctccccctaa ctctccaaac 3481 gtggatcttg ggaagagggc acctacgcct atccgggaaa aggaagtcac catgtgcatg 3541 cgctgccagg agcccttcaa ttccatcacc aaacgcaggc accactgcaa ggcctgtggg 3601 catgtggttt gtgggaaatg ctctgagttc cgagcccgac tcatctatga caacaatcgt 3661 tccaaccgtg tgtgcactga ttgctatgtg gccttgcatg gagcacctgg gagcagtcca 3721 gcctgtagcc agcatacacc ccagcgcagg aggtccatcc tagagaaaca ggcctcggtc 3781 gctgctgaga acagcgtcat ctgcagcttc ctgcactaca tggagaaggg tgggaaagga 3841 tggcacaagg catggtttgt ggttcctgag aatgaaccct tggtgctgta catctatgga 3901 gcccctcagg atgtgaaagc ccagcgcagc ctgcccctca ttggctttga ggtggggcct 3961 cctgaggcag gagagcggcc agacaggagg catgtcttca agatcacaca aagccaccta 4021 agctggtact tcagcccaga gacagaagaa ctccagcggc gctggatggc tgttctcggc 4081 agggcgggcc gtggggacac attctgccca gggcccacac tgtctgagga caaggagatg 4141 gaggagacac cagtggcagc ctcaggagcc actgctgaac ctcctgaagc ctcccagacc 4201 cgagacaaga cctaagaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc 4261 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa 4321 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt 4381 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 4441 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg 4501 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 4561 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg 4621 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 4681 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac 4741 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga 4801 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga 4861 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca 4921 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa 4981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 5041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg 5101 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 5161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 5221 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 5281 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 5341 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 5401 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag 5461 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 5521 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 5581 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 5641 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga 5701 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 5761 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 5821 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 5881 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5941 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 6001 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 6061 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 6121 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 6181 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 6241 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 6301 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 6361 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 6421 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga 6481 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg 6541 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac 6601 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc 6661 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat 6721 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 6781 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 6841 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 6901 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 6961 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 7021 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 7081 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 7141 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 7201 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 7261 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 7321 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 7381 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 7441 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 7501 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 7561 tgtggataac cgtattaccg ccatgcat //