LOCUS       sequence-261611-        6642 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 6642)
  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 6642)
  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-Asef2
FEATURES             Location/Qualifiers
     source          1..6642
                     /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    3430..3551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3467..3486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3521..3540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3558..4013)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3695..3716)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3907..3926
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4040..4144
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4146..4503
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4146..4337
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4169..4189)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4354..4489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4538..5332
                     /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             4538..5332
                     /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(4592..4611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5520..5539)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5564..5611
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5940..6528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6429..6448
                     /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 gatctcgagt tatgacttct gccagccctg aagaccagaa tgctccagtg
     1381 ggctgcccca aaggagcccg gagaaggcgc cccatttccg tgataggtgg ggtcagcttg
     1441 tatgggacca accagacgga ggaactggac aatcttctga cccaaccggc ttccaggccg
     1501 cccatgcctg ctcaccaggt gccaccctac aaggctgtgt cggcccggtt ccggcccttc
     1561 acattctccc agagcacccc cattgggttg gaccgtgtgg gacgccggcg gcagatgaga
     1621 gcatccaacg tttcttcaga tggaggtact gagccctctg ccttagtgga tgacaacggt
     1681 agtgaggagg acttcagcta tgaagacctc tgccaggcca gccctcggta cctgcagccc
     1741 ggcggggagc agctggccat caatgagctg atcagtgatg gcaacgtggt ctgcgcagaa
     1801 gccctgtggg accatgtgac catggatgac caggaactgg gcttcaaagc cggggatgtc
     1861 atccaggttc tggaagcctc caacaaggac tggtggtggg gccgcagtga agataaggaa
     1921 gcctggttcc ccgcgagctt cgtcagattg cgagtgaatc aggaagagct gtcggaaaac
     1981 tccagcagca cccccagtga ggagcaggac gaggaggcca gccagagccg ccacagacac
     2041 tgtgagaaca agcagcagat gcggaccaac gtcatccggg agatcatgga caccgagcgg
     2101 gtgtacatca aacacctcag ggacatctgt gagggctata tccgacagtg ccgcaagcac
     2161 acaggaatgt tcaccgttgc gcagctagcc actatttttg gaaacattga agatatttac
     2221 aaattccaaa gaaagtttct gaaagacctt gagaaacagt acaacaaaga ggagcctcac
     2281 ttaagtgaaa taggatcttg ctttcttcaa aatcaagagg gctttgccat ctattccgag
     2341 tactgcaaca accacccggg cgcctgcctg gagctcgcca acctcatgaa gcagggcaag
     2401 tacagacatt tctttgaagc ctgccgcctg ctgcagcaga tgattgacat cgccatcgac
     2461 gggttcctgc tcacaccagt gcagaagatc tgcaaatacc cgctgcagct ggccgagctg
     2521 ctcaagtata ccacacagga acacggtgat tacagcaaca taaaggcagc atatgaggcc
     2581 atgaagaatg tggcctgtct gatcaacgag cgcaagcgca agctggagag catcgacaag
     2641 atagctcgct ggcaggtgtc tatcgtgggc tgggagggac tggatatctt agaccgaagc
     2701 tcagaattga ttcattctgg ggagctgacc aaaatcacta agcaaggcaa aagccagcag
     2761 cggacgttct tcctgtttga ccaccagctg gtgtcctgca agaaggacct gctgcgcagg
     2821 gacatgctgt actacaaggg ccggctggac atggatgaga tggagcttgt ggacctgggg
     2881 gatgggcgcg acaaggactg caacctcagc gtgaaaaatg ccttcaagct cgtcagtagg
     2941 accacagacg aggtttattt gttttgtgcc aaaaaacaag aagacaaggc gaggtggctg
     3001 caggcctgtg cagatgaaag gaggcgggtg caagaggaca aggagatggg aatggaaatt
     3061 tcagaaaacc agaagaaact tgccatgtta aatgctcaaa aggcaggaca tggaaagtca
     3121 aaaggctaca acaggtgccc cgtggcccca ccgcaccagg gcctgcaccc catccaccag
     3181 cgccacatca ctatgcccac aagcgtcccc cagcagcagg tctttggcct ggcggaaccc
     3241 aagaggaagt cctcgctctt ctggcacacc ttcaacaggc tcaccccctt ccggaaatga
     3301 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     3361 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3421 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3541 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3601 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3661 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3721 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3781 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3841 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3901 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3961 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4021 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4081 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4141 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4261 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4321 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4381 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4441 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4501 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5401 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5461 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5521 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5581 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5641 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5701 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5761 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5821 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5881 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5941 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6001 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6061 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6121 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6181 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6241 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6301 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6361 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6421 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6481 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6541 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6601 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
