LOCUS       sequence-261653-        8869 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 8869)
  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 8869)
  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-PLEKHG2
FEATURES             Location/Qualifiers
     source          1..8869
                     /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"
     polyA_signal    5657..5778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5694..5713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5748..5767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5785..6240)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5922..5943)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6134..6153
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6267..6371
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6373..6730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6373..6564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6396..6416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6581..6716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6765..7559
                     /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             6765..7559
                     /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(6819..6838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7429..7448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7747..7766)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7791..7838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8167..8755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8656..8675
                     /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 gatctatgcc tgagggagcc caaggactga gcctctccaa acctagccca
     1381 agcctcgggt gtggccgaag aggtgaagtg tgtgactgtg gcaccgtgtg tgagactcgg
     1441 acagctcctg cagcccccac catggcctcc ccccgaggtt ctgggagctc cacatccctg
     1501 agcacagtgg gctctgaggg ggatccagcc cctgggccca ctccagcctg ctcagcctcc
     1561 aggccagagc cccttccagg gcctcccatc cgcctacatc tctctccggt ggggatccca
     1621 ggttcagcca gaccctcaag gctggagcgt gtggcccggg agatcgtgga gacagaacgg
     1681 gcctatgtca gggacctccg cagcatcgtg gaggactacc tgggccctct gctggacggc
     1741 ggggtcctgg ggctgagcgt ggagcaggtg ggcacgctgt ttgccaacat tgaggacatc
     1801 tacgagttca gcagcgagct cctggaggac ttggagaaca gcagcagcgc cgggggtatt
     1861 gccgagtgct tcgtgcagag gagcgaggat tttgacatct acacattgta ctgcatgaac
     1921 tacccgagct ccctggccct gctccgggag ctgtcgttgt ctccgccagc agccctgtgg
     1981 ctgcaggagc gccaggccca gcttcgccac tcgctgcccc tgcagagctt cctgctgaaa
     2041 cctgtccagc gcattctcaa gtaccatctg ctgctgcagg aactagggaa gcactgggcg
     2101 gagggcccag gcactggggg tcgcgagatg gtggaggaag ctattgtgtc catgacagcg
     2161 gttgcctggt acatcaacga catgaagcgc aagcaggagc atgcagcgcg cctccaggaa
     2221 gtgcagcggc ggctgggtgg ctggaccgga ccagagctca gtgcttttgg ggaactggtg
     2281 ttggagggcg cgttccgagg aggcggaggg ggtggccccc ggctacgagg gggtgagcgg
     2341 ctgctcttcc tgttctctcg gatgctgctg gtggccaagc gcagggggct ggagtacacc
     2401 tacaaaggcc acatcttctg ctgcaacctg agcgtgagcg agagtccccg agaccctcta
     2461 gggttcaagg tgtctgatct gaccattccc aagcacagac acctgctcca ggccaagaac
     2521 caagaagaga agaggctgtg gattcactgt ctccagcgcc tcttctttga gaaccaccct
     2581 gcctccatcc ctgccaaggc aaagcaagtt ctccttgaaa acagcctgca ttgtgccccc
     2641 aaaagtaagc ctgtcctaga gcccctgaca cccccacttg ggtctcctcg acctcgagat
     2701 gctagaagtt ttacccctgg gcgaaggaac acagctccat ctcctgggcc ctctgtgatt
     2761 cgccgaggcc gcaggcagtc tgagccggtg aaggaccctt atgtcatgtt cccacagaac
     2821 gctaagcctg gattcaagca cgctggcagc gaaggggaac tctaccctcc agaatctcag
     2881 ccaccagttt caggctctgc accccctgag gacctggagg atgctggacc cccaacactg
     2941 gacccctctg ggacctcaat cactgaagaa atcctggaac tgctgaatca gcgaggcctt
     3001 cgagatccag ggccgtccac ccatgacatt cccaagttcc ccggagactc ccaggtgcct
     3061 ggcgacagcg aaaccctcac attccaagcc ctgcccagcc gggactcttc agaagaggag
     3121 gaggaggaag aggaagggct ggagatggat gaacgggggc cttccccact ccacgtcctg
     3181 gaagggctcg aaagttccat tgcagctgaa atgcccagca ttccctgcct taccaaaatt
     3241 cctgacgtgc ccaaccttcc tgaaattccc agccgctgtg aaattcccga aggttctcgc
     3301 cttcctagtc tctctgacat ttccgatgtt tttgagatgc cctgccttcc agccatacct
     3361 agtgtcccca acacccccag tctgtctagc actcccaccc tctcctgtga ctcctggctc
     3421 caagggcctc tgcaggaacc agctgaggct ccagccacca ggagagaact gttttctggg
     3481 agcaatcctg ggaaactggg agagccgcct tcaggaggca aggcagggcc agaggaggat
     3541 gaagaagggg tatcattcac agacttccag ccccaggatg tcacccaaca tcagggattc
     3601 ccagatgagc tggcattccg ctcttgctca gaaatccgga gcgcctggca ggcattggaa
     3661 cagggacagc tggcccggcc aggcttccca gagccactgc tgatcctgga ggattcggat
     3721 ctgggtggag acagcgggag cgggaaggca ggagccccga gttcagaaag gacggcgtcc
     3781 cgagtgcgag agctggcccg gctttacagc gagcggatcc agcagatgca gcgggcggag
     3841 actcgggcat cagccaatgc cccgcgccgc cggcctcggg ttctggccca accccagcca
     3901 tccccctgtc tgccccagga gcaggcagag ccagggctcc tgcctgcctt tggacacgtg
     3961 ctggtatgtg agctggcctt cccactgaca tgtgcccagg agtctgtccc cctgggtcct
     4021 gctgtctggg ttcaagctgc catacctttg tcaaagcagg gaggcagccc ggatggccag
     4081 ggtctacatg tttccaattt gcctaagcaa gaccttccgg gcatccacgt ttcagctgct
     4141 acccttttgc ctgagcaagg aggttcccgg catgtccagg ctccagccgc cacacctttg
     4201 cccaagcaag aaggccccct gcacctccag gtgccggctc ttacaacttt ctctgatcaa
     4261 ggccacccag aaatccaagt tccagccacc actcctttgc ctgagcataa aagtcacatg
     4321 gttataccag ctccatccac cgccttttgt cctgagcagg gacactgtgc ggacatccac
     4381 gttcccacca ctccagcttt gcccaaggag atttgttctg atttcacagt ttcagtcacc
     4441 acccctgtgc ccaagcaaga aggtcaccta gacagcgaga gcccaaccaa tatcccactg
     4501 acaaagcaag gaggttccag ggatgttcag ggcccagacc ctgtctgcag tcaacccatc
     4561 cagcctttgt cttggcatgg aagcagcctg gatccccagg gcccaggcga caccctacca
     4621 cccttgccat gtcacctccc agaccttcag attccaggta cctcaccttt gcctgcacat
     4681 ggaagccacc tggaccatcg gatcccagcc aacgccccac tgtctttgtc ccaggagctc
     4741 ccagacactc aggttccagc taccacacct ttgcccctgc cacaagtcct cacagacatc
     4801 tgggtccaag ccctcccaac ttcacccaag cagggaagcc tcccagacat ccagggtcca
     4861 gcggctgcac ctccacttcc ggagccaagc cttacagata cacaggtcca aaaactcaca
     4921 ccttcgttgg agcagaagag cctcatagat gcccatgttc cagctgccac acctttacct
     4981 gagagaggag gctctctaga cattcagggc ctctcaccca ccccagttca gaccaccatg
     5041 gttttgtcca aaccaggagg ctccttagcc tctcacgttg ccaggttgga gtcttcagac
     5101 ttgacgccac ctcatagtcc cccaccttcc agccgtcagc tcctgggccc caatgcagct
     5161 gccctctcca gatacctggc agcctcatat atcagccaaa gcctggctcg gcggcagggg
     5221 cctgggggag gggcccccgc agcctcccgg ggctcctggt cctctgctcc cacgtcacgg
     5281 gcatcttcgc cgccccccca gccccagcca ccacctcccg cagccaggcg gctcagctat
     5341 gccacgacgg ttaacatcca cgtgggcggg ggtgggcggc tgcggccagc caaggcccag
     5401 gtccggttga accaccctgc tctcttggcc tccacacagg aatctatggg ccttcacagg
     5461 gcccaggggg ctcctgatgc ccccttccac atgtgagaat tctgcagtcg acggtaccgc
     5521 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     5581 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5641 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5701 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5761 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5821 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5881 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5941 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6001 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6061 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6121 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6181 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6241 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6301 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6361 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6481 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6541 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6601 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6661 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6721 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6781 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6841 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6901 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6961 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     7021 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     7081 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     7141 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     7201 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7261 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     7321 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7381 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7441 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7501 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7561 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7621 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7681 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7741 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7801 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7861 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7921 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7981 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     8041 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8101 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8161 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8221 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8281 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8341 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8401 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8461 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8521 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8581 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8641 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8701 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8761 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8821 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
