LOCUS       sequence-261655-        8868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RhoGEF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8868)
  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 8868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Small 
            GTPases. 2019 Aug 30:1-15. doi: 10.1080/21541248.2019.1658509."
COMMENT     Sequence Label: mCherry-PLEKHG1
FEATURES             Location/Qualifiers
     source          1..8868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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"
     RBS             1476..1484
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5656..5777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5693..5712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5747..5766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5784..6239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5921..5942)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6133..6152
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6266..6370
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6372..6729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6372..6563
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6395..6415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6580..6715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6764..7558
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6764..7558
                     /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(6818..6837)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7428..7447
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7746..7765)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7790..7837
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8166..8754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8655..8674
                     /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 gatctcgagc cgagctctct gatagtgacc gacccgtcag cttcggttcc
     1381 acatcatcct cggcctcttc ccgcgacagc catggttcct tcggcagcag aatgacctta
     1441 gtttcaaata gccacatggg cttgtttaac caggataagg aggtaggggc cataaaactg
     1501 gagctgattc ctgccaggcc gttttccagc agcgagctgc agagggacaa ccccgccacg
     1561 gggcaacaga acgcggatga gggcagcgaa aggccaccca gagcgcagtg gagagtggac
     1621 tcaaacgggg cacccaagac gatcgcagac tcggccacga gccccaagct cctctatgtg
     1681 gatagagttg ttcaggaaat tctggaaacc gaaaggactt atgtacaaga tttaaaaagc
     1741 atcgtagagg attaccttga ctgcatcagg gaccaaacaa aacttcccct ggggaccgaa
     1801 gaaagatcag ccctttttgg aaacatacag gatatctacc acttcaatag tgaacttctg
     1861 caagatttgg aaaactgtga aaatgatcct gtggccatag cagagtgttt tgtgtccaag
     1921 agtgaagagt tccacattta tacccagtat tgcactaact atccaaggtc cgtggctgtg
     1981 ctaacagagt gtatgaggaa caagatactg gccaaattct tcagggagcg tcaggaaact
     2041 ctgaaacact cgctgcctct ggggtcctat ctcttgaaac cagttcagcg gattctcaag
     2101 tatcatctcc ttctgcatga aatagaaaac caccttgata aggacacaga aggctatgat
     2161 gtggtgcttg atgctataga cacaatgcag cgagtcgcct ggcatatcaa tgacatgaag
     2221 cggaaacacg agcacgcggt ccggttacag gagatacaga gtttgctcac taactggaag
     2281 gggccagacc tgaccagcta cggggaactg gtgcttgagg gaaccttccg catccagcga
     2341 gccaagaatg agcggacgct cttcctcttc gacaagctgc tgctcatcac gaagaagaga
     2401 gatgacacgt ttacatacaa agctcacatc ctgtgtggca acctcatgct tgtggaggtg
     2461 attccaaaag agccgctcag cttcagcgtc ttccactaca agaatcccaa gctgcagcac
     2521 acagtccagg ccaaatccca gcaagacaaa cgcctctggg ttctgcacct aaagagactg
     2581 attctggaga accatgcagc caagattcca gctaaggcca agcaagcaat ccttgaaatg
     2641 gatgccattc atcatccagg cttctgttac agtcctgagg gagggacgaa agcactgttc
     2701 ggctctaaag aaggttctgc tccatatcgg ctgagaagaa aatctgaacc ttcctcacgg
     2761 tcacataaag ttttgaagac cagtgaaaca gcacaagaca tccaaaaggt ttctagagag
     2821 gaaggatctc cccagctgtc ttcagcaagg ccgtctcctg cccagagaaa tagtcagcct
     2881 agcagttcta ccatgatcag cgtgcttcga gcgggtggag ctctcagaaa catctggacc
     2941 gatcaccaga tcaggcaagc cttgtttccc agccgacggt ccccgcagga gaatgaggat
     3001 gatgaagatg attatcagat gttcgtgccg tcattttcct cctcagatct gaactctacc
     3061 agactatgtg aagatagcac ttctagtcgc ccttgcagct ggcatatggg acagatggag
     3121 tccacagaga cctccagctc tggtcacaga attgtcaggc gggccagcag tgctggggag
     3181 agcaacacat gccctcctga aataggaact agtgacagaa ctagggaact gcagaacagc
     3241 cccaaaacag aagggcagga ggagatgact ccctttgggt catccataga gttgactatt
     3301 gatgacatag accatgtcta tgataacatc agttatgagg acttaaaact aatggttgct
     3361 aagcgggaag aagctgaatc cactccctct aaatcagcca gggactccgt tcgccccaag
     3421 agcaccccag agttagcctt cacaaagagg caagctggcc acagtaaggg ctctctttac
     3481 gcacaaacag atggcaccct ctcaggtgga gaggcatcca gccaaagcac gcatgaactc
     3541 caagcggttg aggagaacat ctatgacacc atagggctcc cagatcctcc gtcgctgggt
     3601 tttaagtgca gcagcctaaa gcgtgcaaag cggagcacct ttttgggtct ggaggccgac
     3661 ttcgtgtgct gtgacagcct gaggccattt gtttcccaag acagcctcca gctcagtgag
     3721 gacgaagccc cttaccatca ggccaccccc gatcatggtt atctgagttt gctgtatgac
     3781 tctcccagtg gtaacttgtc tatgcctcat aagcctgtat ctgataaact gtccgaagaa
     3841 gtagatgaaa tctggaatga cctggaaaat tacatcaaga aaaatgaaga caaggccaga
     3901 gaccgtctcc tggcagcgtt tcctgtgagc aaggatgatg tgccagacag gctgcacgca
     3961 gagagcaccc ctgagctgag ccgggacgtg gggcgctctg tgtccacgct gtccctgcct
     4021 gagagccagg ctctcctcac gcccgtgaag agcagggctg gcagagccag ccgcgccaac
     4081 tgcccctttg aggaagacct gatttctaaa gaaggctcct ttatgagcct taaccggctt
     4141 tctctggcta gtgaaatgcc cctcatggac aatccctacg acctggccaa cagtggcctg
     4201 tctcaaacag acccagaaaa ccctgacctg gggatggagg ccacagataa gacaaaaagc
     4261 agggtgttta tgatggctcg gcagtacagt cagaagatta agaaggcgaa tcaactttta
     4321 aaagtgaaaa gtctggagct ggagcagccg ccggccagtc agcatcagaa atccatgcac
     4381 aaagacctgg ctgccatctt ggaagagaag aagcaaggag ggcccgccat tggtgccagg
     4441 attgctgagt attcccagtt gtatgaccag attgtattca gggagtctcc cttgaaaatt
     4501 cagaaggatg gctgggccag ccctcaagaa tcctccctcc tgaggtctgt gtcaccttcc
     4561 caggtccacc atggtagtgg agactggctt ctgcattcaa cctatagtaa tggagagtta
     4621 gcagatttct gtctcccacc agagcaagac ttgaggtcaa gatatcccac gtttgagatc
     4681 aatacaaaaa gtactcccag gcaattgtcc gcagcttgct ctgtgccttc tcttcaaacc
     4741 tctgaccctc tgccaggctc tgtgcagaga tgcagcgtgg tagtaagtca gcccaacaaa
     4801 gagaactggt gtcaggacca tctttacaac tccttgggtc ggaaagggat cagcgctaaa
     4861 tctcagcctt atcacaggtc ccagtcatct tcctccgtct tgatcaacaa atcaatggat
     4921 tccatcaact accctagtga tgtgggaaag cagcagctgc tgtctttaca cagaagttca
     4981 aggtgtgaga gtcaccagga cttgctgcca gatattgctg actcgcatca acagggcact
     5041 gaaaaactct cagatctcac actccaagac tcacagaaag ttgtggtggt caatagaaat
     5101 ttacccttaa atgcccaaat tgcaacacag aattattttt ccaatttcaa agagactgat
     5161 ggagatgaag atgactatgt ggaaatcaag tcagaagaag atgagtcgga gttggagcta
     5221 tctcacaatc gtagaaggaa atctgactca aagtttgtgg atgctgactt ttctgataat
     5281 gtctgcagcg gcaacacatt gcattctttg aatagtccgc gcactccaaa aagccggtta
     5341 acagcaaact tggcctttca ccatatctga caccatataa tgattctgac aaactgaatg
     5401 actacctttg gagggggcca tctcccaatc aacaaaatat tgtccagtct ctaagggaaa
     5461 aatttcagtg tctcagttca agcagctttg cttaagaatt ctgcagtcga cggtaccgcg
     5521 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     5581 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5641 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5701 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5761 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     5821 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5881 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5941 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6001 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6061 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6121 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6181 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6241 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6301 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6361 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     6421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6721 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6781 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6841 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6901 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     6961 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     7021 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     7081 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     7141 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     7201 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     7261 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     7321 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7381 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7441 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7501 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     7561 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     7621 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     7681 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     7741 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     7801 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     7861 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     7921 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     7981 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     8041 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     8101 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     8161 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8221 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8281 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     8341 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8401 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8461 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8521 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8581 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8641 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8701 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8761 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8821 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
