LOCUS       sequence-261591-        7335 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 7335)
  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 7335)
  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-Vav2
FEATURES             Location/Qualifiers
     source          1..7335
                     /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    4123..4244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4160..4179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4214..4233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4251..4706)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4388..4409)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4600..4619
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4733..4837
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4839..5196
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4839..5030
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4862..4882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5047..5182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5231..6025
                     /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             5231..6025
                     /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(5285..5304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5895..5914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6213..6232)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6257..6304
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6633..7221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7122..7141
                     /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 catggagcag tggcgacagt gcggccgctg gctcatcgat
     1381 tgcaaggtcc tgccgcccaa ccaccgggtg gtgtggccct cggccgtggt cttcgacctg
     1441 gcgcaggcgc tgcgcgacgg ggtccttctg tgccagctgc tgcacaacct ctcccccggc
     1501 tccatcgacc tcaaggacat caacttccgg ccgcagatgt cccagtttct gtgtttgaag
     1561 aacatacgca ccttcctgaa agtctgccac gataaatttg gattaaggaa cagcgagctg
     1621 tttgacccct ttgacctctt cgatgtgcga gactttggaa aggtcatctc cgcggtgtcg
     1681 aggctctccc tgcacagcat cgcgcagaac aaagggatca ggccttttcc ctcagaggag
     1741 accacagaga atgacgatga cgtctaccgc agcctggagg agctggccga cgagcatgac
     1801 ctgggggagg acatctacga ctgcgtcccg tgtgaggatg gaggggacga catctacgag
     1861 gacatcatca aggtggaggt gcagcagccc atgattagat acatgcagaa aatgggcatg
     1921 actgaagatg acaagaggaa ctgctgcctg ctggagatcc aggagaccga ggccaagtac
     1981 taccgcaccc tggaggacat tgagaagaac tacatgagcc ccctgcggct ggtgctgagc
     2041 ccggcggaca tggcagctgt cttcattaac ctggaggacc tgatcaaggt gcatcacagc
     2101 ttcctgaggg ccatcgacgt gtccgtgatg gtggggggca gcacgctggc caaggtcttc
     2161 ctcgatttca aggaaaggct cctgatctac ggggagtact gcagccacat ggagcacgcc
     2221 cagaacacac tgaaccagct cctggccagc cgggaggact tcaggcagaa agtcgaggag
     2281 tgcacactga aggtccagga tggaaaattt aagctgcaag acctgctggt ggtccccatg
     2341 cagagggtgc tcaaatacca cctgctcttg aaggagcttc tgagccattc tgcggaacgg
     2401 cctgagaggc agcagctcaa agaagcactg gaagccatgc aggacttggc gatgtacatc
     2461 aatgaagtta aacgggacaa ggagaccttg aggaaaatca gcgaatttca gagttctata
     2521 gaaaatttgc aagtgaaact ggaggaattt ggaagaccaa agattgacgg ggaactgaaa
     2581 gtccggtcca tagtcaacca caccaagcag gacaggtact tgttcctgtt tgacaaggtg
     2641 gtcatcgtct gcaagcggaa gggctacagc tacgagctca aggagatcat cgagctgctg
     2701 ttccacaaga tgaccgacga ccccatgaac aacaaggacg tcaagaagtc tcacgggaaa
     2761 atgtggtcct acggcttcta cctaattcac cttcaaggaa agcagggctt ccagtttttc
     2821 tgcaaaacag aagatatgaa gaggaagtgg atggagcagt ttgagatggc catgtcaaac
     2881 atcaagccag acaaagccaa tgccaaccac cacagtttcc agatgtacac gtttgacaag
     2941 accaccaact gcaaagcctg caaaatgttc ctcaggggca ccttctacca gggatacatg
     3001 tgtaccaagt gtggcgtcgg ggcacacaag gagtgcctgg aagtgatacc tccctgcaag
     3061 ttcacttctc ctgcagatct ggacgcctcc ggagcgggac caggtcccaa gatggtggcc
     3121 gtgcagaatt accatggcaa cccagcccct cccgggaagc ctgtgctgac cttccagacc
     3181 ggcgacgtgc ttgagctgct gaggggcgac cctgagtctc cgtggtggga gggtcgtctg
     3241 gtacaaacca ggaagtcagg gtatttcccc agctcatctg tgaagccctg ccctgtggat
     3301 ggaaggccgc ccatcagccg gccgccatcc cgggagatcg actacactgc atacccctgg
     3361 tttgcaggta acatggagag gcagcagacg gacaacctgc tcaagtccca cgccagtggg
     3421 acctacctga tcagggagcg gcctgccgag gctgagcgct ttgcaataag catcaagttc
     3481 aatgatgagg tgaagcacat caaggtggtg gagaaggaca actggatcca catcacagag
     3541 gccaagaaat tcgacagcct cctggagttg gtggagtact accagtgcca ctcactgaag
     3601 gagagcttca agcagctgga caccacactc aagtacccct acaagtcccg ggaacgttcg
     3661 gcctccagag cctccagccg gtccccagct tcctgtgctt cctacaactt ttcttttctc
     3721 agtcctcagg gcctcagctt tgcttctcag ggcccctccg ctcccttctg gtcagtgttc
     3781 acgccccgcg tcatcggcac agctgtggcc aggtataact ttgccgcccg agatatgagg
     3841 gagctttcgc tgcgggaggg tgacgtggtg aggatctaca gccgcattgg cggagaccag
     3901 ggctggtgga agggcgagac caacggacgg attggctggt ttccttcaac gtacgtagaa
     3961 gaggagggca tccagtaagg taccgcgggc ccgggatcca ccggatctag ataactgatc
     4021 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4081 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4141 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4201 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     4261 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4321 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4381 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4441 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4501 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4561 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4621 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4681 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4741 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4801 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4861 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4921 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4981 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5041 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5101 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5161 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5221 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5281 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5341 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5401 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5461 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5521 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5581 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5641 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5701 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5761 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5821 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5881 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5941 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6001 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6061 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6121 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6181 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6241 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6301 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6361 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6421 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6481 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6541 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6601 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6661 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6721 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6781 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6841 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6901 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6961 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7021 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7081 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7141 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7201 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7261 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7321 attaccgcca tgcat
//
