LOCUS       sequence-261661-        9097 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 9097)
  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 9097)
  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-FGD5
FEATURES             Location/Qualifiers
     source          1..9097
                     /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    5885..6006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5922..5941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5976..5995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6013..6468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6150..6171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6362..6381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6495..6599
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6601..6958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6601..6792
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6624..6644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6809..6944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6993..7787
                     /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             6993..7787
                     /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(7047..7066)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7657..7676
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7975..7994)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8019..8066
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8395..8983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8884..8903
                     /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 gatctatgtt caggggtccg aagcccccca ttgcccccaa gcccaggctg
     1381 actgccccaa acgagtggag agccagtgtg tacctgaatg acagcttgaa caaatgcagc
     1441 aacgggcggc tgccctgtgt agacaggggg cttgatgagg ggccccggtc catcccaaag
     1501 tgctctgagt cggagaccga cgaggattac atcgtggtcc ccagggttcc gctgagggag
     1561 gatgaaccca aggacgaggg cagtgtgggg aacaaagccc tggtgtctcc cgagtcctct
     1621 gcggaagagg aagaggagcg tgaagaggga ggcgaggcat gtggcctgga gggtacagga
     1681 gctggtgagg attcagtggc ccctgctgct ccgggtgcag gagcgctgag cagggagggt
     1741 gaggaaggca cagaccttgc tcttgaggat gaaggggagg gctgcgctga tgagccaggg
     1801 acactggagc aggtgtccag aagtgaggag gaagagaagc tagtgcagcc acacagggag
     1861 tgcagcctgg aggacagtgg gccttgggct ggagaggggg tcttccagag cgacctcctc
     1921 ctgcctcaca tccatggaga ggaccaggag ccccccgaca cccccgggga ggcagaggag
     1981 gatgatgagg aaggctgtgc cagcacagac ccagcagggg cagatgaggg ttcgggtcct
     2041 gacaggccca cggaggacat gggacaggat gctgaggaca ccagtgagga gccccctgag
     2101 aaggaggagc tggccggggt ccaggaggca gagacagcca cagactgccc tgaagttctt
     2161 gaggagggat gtgaagaggc cacgggtgtc acaggtgggg aacaggttga cctcagtgaa
     2221 ccacctgacc acgagaagaa aaccaaccaa gaagtggcag ccgccaccct ggaggaccat
     2281 gcacaggatg agtccgccga ggagagctgc cagattgtcc cttttgagaa tgactgcatg
     2341 gaggacttcg tgacttccct cacaggaagc ccctatgagt tcttcccaac tgagagcacc
     2401 tctttttgca gcgagagctg ttctcctctt tctgaatcag cgaaaggttt agaatcagag
     2461 caggcaccaa agctggggct gcgtgcggag gagaacccca tggtgggggc tttgtgtggc
     2521 cagtgtggct ccctacaggg tggagcggcc gagggtcccg cagcccctga tgtggtggtc
     2581 gtgctggagg aggaggcctt ggatgatgca ctggccaacc cctatgtgat gggagtgggc
     2641 ctgcccggtc aggcggcccc tggagaagga gggcaggctg catcggacgc cctgggtggt
     2701 tatggctcga aagaagaatt gaactgtgag gcagagggtg gcctggttcc cgcggacagg
     2761 aagaacacca gcacgagggt ccggccccac tctgggaagg tggccggcta tgtcccagaa
     2821 accgtccctg aagaaaccgg acctgaggcg ggctcgtcag cccctggcat tggaggtgcc
     2881 gcagaggagg tgggaaagac gcttttgtca ttggagggga agcccttgga agccagcagg
     2941 gccttgccag caaagcccag ggcctttact ttataccctc ggtcgttctc cgtggaaggc
     3001 cgagagattc cagtgtccgt gtaccaggag cctgaggggt cagggttgga tgaccacagg
     3061 ataaagagga aagaggacaa tctctctctg tcgtgtgtaa ttggctcctc tgggagtttc
     3121 tcccagagaa accaccttcc gtccagcggc acctccacgc cttcttccat ggtcgacatc
     3181 ccacctcctt tcgacctggc ctgcatcacc aagaagccca tcacaaagag ctctccctca
     3241 ctcctgatcg agagcgactc cccggacaag tacaagaaga agaagtcatc ctttaagcgc
     3301 ttcctggcac tgacgtttaa gaagaagacg gagaacaaat tgcatgtgga tgtgaacgtg
     3361 tcttcctcta ggtcctcttc agagtccagc taccacgggc cttccaggat tctggaagtt
     3421 gaccggagaa gcctcagcaa ctcccctcag cttaagtctc ggactgggaa gctccgggct
     3481 tctgaatccc cctcctccct catcttttat agagatggca agaggaaagg tgtccccttc
     3541 agcaggacgg tgtccagagt ggagtccttt gaagaccgct cccggccgcc cttcctgccc
     3601 ttgccactga ccaagccacg gtccatctcc ttccccagcg ctgacacttc agactatgag
     3661 aacattccag ccatgaactc ggactatgag aatatccaga ttccaccccg gagacctgcc
     3721 agggctggcg cgttcacgaa gctgtttgaa gatcagagca gagccctgtc cacagcaaac
     3781 gaaaatgatg gctacgtgga catgagcagc ttcaacgcct ttgagagcaa acagcagagt
     3841 gcagaccagg acgcagaaag cgcctacaca gagccctaca aagtctgtcc catctcgtcg
     3901 gcagccccca aagaggacct tacgtcggat gaagagcaga gaagctcgga ggaggaggac
     3961 agtgcttcaa gagaccccag tgtcacccac aaggtggaag gacagtccag agcccttgtc
     4021 atcgcacagg aactgctatc ttcagagaaa gcatacgtgg agatgctcca gcacttaaat
     4081 ctggatttcc atggagctgt catgagggcc ttggatgaca tggaccatga aggcagagac
     4141 acattggccc gggaggagct gaggcagggc ctgagtgaac tcccagccat ccacgacctt
     4201 catcaaggca tcctggagga gctggaggaa aggctgtcaa attgggagag ccagcagaag
     4261 gtagctgacg tcttccttgc ccgggagcag gggtttgatc accacgccac tcacatcctg
     4321 cagttcgaca ggtacctagg tctgctcagt gagaattgcc tccactctcc ccggctggca
     4381 gctgctgtcc gtgaatttga gcagagtgta caaggaggca gccagactgc gaagcatcgg
     4441 ctgctgcggg tggttcaacg cctcttccag taccaagtgc tcctcacaga ctatttaaac
     4501 aacctttgtc cggactccgc cgagtacgac aacacacagg gtgcactgag cctcatctcc
     4561 aaagtcacag accgtgccaa cgacagcatg gagcaagggg aaaacctgca gaagctggtc
     4621 cacattgagc acagcgtccg gggccaaggg gatctcctcc agccaggaag ggagtttctg
     4681 aaggaaggga cgctgatgaa agtaacaggg aaaaacagac ggccccggca cctatttctg
     4741 atgaacgatg tgctcctgta cacctatccc cagaaggatg ggaagtaccg gctgaagaac
     4801 acattggctg tggccaacat gaaggtcagc cgccctgtga tggagaaagt gccctacgct
     4861 ctaaagattg agacttccga gtcctgcctg atgctgtctg cgagctcctg tgcagagagg
     4921 gacgagtggt atggctgtct gagcagagcc ctccctgagg actacaaggc ccaggcgctg
     4981 gctgcattcc accatagcgt ggagatacga gagaggctgg gggttagcct tggggagagg
     5041 ccccccaccc tggtgcctgt cacacacgtc atgatgtgca tgaactgcgg ctgcgacttc
     5101 tccctcaccc tgcggcgtca tcactgtcac gcctgtggca agatcgtgtg ccggaactgt
     5161 tcgcggaaca agtacccgct gaagtacctg aaggacagga tggccaaggt ctgcgacggc
     5221 tgcttcgggg agctgaagaa gcggggcagg gctgtcccgg gcctgatgag agagcggcct
     5281 gtgagcatga gcttcccgct gtcttcaccc cgcttctcgg gcagtgcctt ttcatccgtc
     5341 ttccagagca ttaacccctc gaccttcaag aagcagaaga aagtcccttc agccctgaca
     5401 gaggtggctg cctctggaga gggctctgcc atcagtggct atctcagccg gtgtaagagg
     5461 ggcaagcggc actggaagaa gctctggttt gtcatcaaag gcaaagttct ctacacctac
     5521 atggccagtg aggacaaagt ggccttggag agtatgcctc tgctaggctt caccattgct
     5581 ccagaaaagg aagagggcag cagtgaagta ggacctattt ttcaccttta ccacaagaaa
     5641 accctatttt atagcttcaa agcagaagat accaattcag ctcagaggtg gatcgaggcc
     5701 atggaagatg cgagtgtgtt ataagaattc tgcagtcgac ggtaccgcgg gcccgggatc
     5761 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     5821 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     5881 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5941 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6001 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     6061 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6121 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6181 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6241 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6301 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6361 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6421 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     6481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     6601 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     6721 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6781 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6841 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6901 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     6961 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     7021 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     7081 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     7141 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     7201 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     7261 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     7321 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     7381 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     7441 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     7501 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     7561 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     7621 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     7681 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     7741 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     7801 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     7861 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     7921 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     7981 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     8041 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     8101 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     8161 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     8221 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     8281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     9061 ccctgattct gtggataacc gtattaccgc catgcat
//
