LOCUS       sequence-266099-        8271 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 8271)
  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 8271)
  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-PLEKHG4
FEATURES             Location/Qualifiers
     source          1..8271
                     /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    5059..5180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5096..5115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5150..5169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5187..5642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5324..5345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5536..5555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5669..5773
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5775..6132
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5775..5966
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5798..5818)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5983..6118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6167..6961
                     /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             6167..6961
                     /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(6221..6240)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6831..6850
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7149..7168)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7193..7240
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7569..8157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8058..8077
                     /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 cgaaaggccc ctggagaatg gggatgagtc cccagactct
     1381 cagggccatg ccaccgactg gagatttgct gtgtgcagtt tcagggatgc ctgggaagag
     1441 gaggaacctg ctccccagat gcacgttaag gacccaggtc ctccaagacc accagccggg
     1501 gccacccagg atgaggagct acagggcagc cccttgtcca ggaaattcca gttaccccca
     1561 gctgcagatg agtcggggga tgcccagagg ggcacagtag aaagctcctc agtcctgtca
     1621 gaagggccag gcccctctgg agtggagagt ctcctatgcc ccatgtcctc ccacctcagc
     1681 ttggcacagg gtgagagtga caccccaggg gtagggttgg taggggaccc aggtccaagc
     1741 agggcgatgc catctggctt gagccctggg gcattggaca gcgaccctgt gggccttgga
     1801 gaccctttat cagaaatatc aaagctgctg gaggcagccc ccagtggatc cggcctccct
     1861 aagcctgctg actgcctcct ggcccaagac ctctgttggg agctgctggc cagtggtatg
     1921 gccaccttgc cagggactcg ggatgtccaa ggccgggcag tgctgcttct gtgtgcccac
     1981 agcccagcct ggcttcagtc tgagtgcagc agccaggaac tcatccgcct cctgctgtac
     2041 ctgcgaagca tccccaggcc cgaagtacag gcactgggac tgacagtgct agttgatgcc
     2101 cgaatttgtg ctccaagttc ttccctcttc tctgggctca gccaactaca agaagcagcc
     2161 ccaggggccg tgtaccaggt gctgctagtg ggaagcacgc tgctgaagga agtgccttcc
     2221 gggctgcagc tggagcagtt gccttctcag agcctgctga cccacatccc aacggcgggg
     2281 ctgcccactt cgctaggagg aggcctgcct tactgccacc aggcctggct ggatttccga
     2341 aggcggctgg aagctctact acagaactgc caggcagctt gtgccctgct ccagggggcc
     2401 atcgaaagtg tgaaggctgt gccccagccc atggagcctg gggaggtcgg tcagctgcta
     2461 cagcagacag aggtcctgat gcagcaggtg ctagactcgc catggctggc atggctacaa
     2521 tgccaggggg gccgggagct gacatggctg aagcaagagg tcccagaggt gaccctgagc
     2581 ccagactaca ggacggcaat ggacaaggct gacgagctat atgaccgggt ggatggattg
     2641 ctgcaccaac tgaccctgca gagcaaccag cgaatacagg ccctagagtt ggtccaaaca
     2701 ctggaggccc gggaaagcgg actgcaccag attgaagtgt ggctgcagca ggtgggctgg
     2761 ccagcactgg aggaggctgg ggagccctcg ctggacatgc tgctccaggc ccaaggctct
     2821 tttcaggagc tgtaccaggt tgcccaggag caggtcaggc aaggggagaa gtttctgcag
     2881 ccgctgactg gctgggaggc ggctgaactg gacccccctg gggcacgctt tctggccctg
     2941 cgagcccagc tgactgaatt ctctagggct ttggcccagc ggtgccagcg gctggcggat
     3001 gctgagaggc tgtttcagct cttcagggag gccttgacgt gggctgagga ggggcagcga
     3061 gtgttggcag agctggagca ggaacgcccg ggggttgtgt tgcagcagct gcagctgcac
     3121 tggaccaggc accctgactt gcctcctgcc cacttccgaa agatgtgggc tctggccacg
     3181 gggctgggct cagaggccat ccgccaggag tgccgctggg cctgggcgcg gtgccaggac
     3241 acctggctgg ccctggacca aaagcttgag gcttcactga agctaccacc ggtgggcagc
     3301 acagctagcc tgtgtgtcag ccaggtcccc gctgcacctg cccaccctcc cctgaggaag
     3361 gcctacagct tcgatcggaa tctggggcag agtctcagtg aacctgcctg ccactgccac
     3421 catgcggcca ctattgctgc ctgccgcaga ccagaggctg gaggaggtgc cctgccccag
     3481 gcatccccta ctgtgcctcc accaggcagc tctgacccca ggagcctcaa caggctacag
     3541 ctggtgctgg cagagatggt ggccacggag cgggagtatg tccgggctct agagtacact
     3601 atggagaact atttccccga gctggatcgc cccgatgtgc cccagggcct ccgcggtcag
     3661 cgtgcccacc tctttggcaa cctggagaag ctgcgggact tccactgcca cttcttcctg
     3721 cgtgagctgg aggcttgcac ccggcaccca ccacgagtgg cctatgcctt cctgcgccat
     3781 agggtgcagt ttgggatgta cgcgctctac agcaagaata agcctcgctc cgatgccctg
     3841 atgtcaagct atgggcacac cttcttcaag gacaagcagc aagcactggg ggaccacctg
     3901 gacctggcct cctacctgct aaagcccatc cagcgcatgg gcaagtacgc actgctgctg
     3961 caggagctgg cacgggcctg cgggggcccc acgcaggagc tcagtgcgct gcgggaggcc
     4021 cagagccttg tgcacttcca gctgcggcac ggaaacgacc tgctggccat ggacgccatc
     4081 cagggctgtg atgttaacct caaggaacag gggcagctgg tgcgacagga tgagtttgtg
     4141 gtgcgcactg ggcgccacaa gtccgtgcgc cgcatcttcc tttttgagga gctgctgctc
     4201 ttcagcaagc ctcgccatgg gcccacaggg gttgacacat ttgcctacaa gcgctccttc
     4261 aagatggcag accttggtct cactgagtgc tgtgggaaca gcaacctgcg cttcgagatc
     4321 tggttccgcc gccgcaaggc cagggacacc tttgtgctgc aggcctccag cctggctatc
     4381 aagcaggcct ggacagctga catctcccac ctgctttgga ggcaggccgt ccacaacaag
     4441 gaggtgcgca tggctgagat ggtgtccatg ggtgtgggga acaaggcctt ccgagacatt
     4501 gctcccagcg aggaagccat caacgaccgc accgtcaact atgtcctgaa gtgccgagaa
     4561 gttcgctctc gggcgtccat tgccgtagcc ccgtttgacc atgacagcct ctacctgggg
     4621 gcctcgaact cccttcctgg agaccctgcc tcttgctctg ttctggggtc cctcaacctg
     4681 cacctgtaca gagacccagc tcttctgggt ctccgctgtc ccctgtatcc cagcttccca
     4741 gaggaagcag cactggaggc tgaggcagag ctgggcggcc agccctcttt gactgctgag
     4801 gactcagaga tctcgtccca atgcccatca gccagtggct ccagtggctc tgacagcagc
     4861 tgtgtgtcag ggcaggccct gggtaggggc ctggaggact taccctgtgt ctaaggtacc
     4921 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     4981 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5041 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5161 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     5221 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5281 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5341 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5401 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5461 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5521 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5581 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5641 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5701 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5761 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     5821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5881 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5941 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6001 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6061 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6121 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     6181 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     6241 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     6301 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     6361 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     6421 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     6481 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     6541 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     6601 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     6661 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     6721 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     6781 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     6841 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6901 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6961 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     7021 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     7081 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     7141 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     7201 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     7261 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     7321 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     7381 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     7441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7681 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     8041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
