LOCUS       sequence-261602-        9429 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 9429)
  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 9429)
  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-TUBA
FEATURES             Location/Qualifiers
     source          1..9429
                     /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    6217..6338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6254..6273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6308..6327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6345..6800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6482..6503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6694..6713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6827..6931
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6933..7290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6933..7124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6956..6976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7141..7276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7325..8119
                     /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             7325..8119
                     /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(7379..7398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7989..8008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8307..8326)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8351..8398
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8727..9315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9216..9235
                     /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 tcaagcttcg aattctgcag tcgacatgga ggctggctca
     1381 gtggttcgag ccatttttga cttctgccct agcgtatcag aagaactgcc gctctttgtg
     1441 ggagatatta ttgaggtgct ggcagtggtg gatgaatttt ggcttctagg aaagaaggag
     1501 gatgttacag gacaattccc cagcagtttt gtggaaattg tgaccattcc cagtctaaaa
     1561 gagggagaga ggctgtttgt gtgcatttgt gaattcacat cccaagagtt ggataatctt
     1621 cccctccatc gaggtgacct ggtgattctc gatggcattc ccactgcagg ctggctgcag
     1681 ggccgaagct gctggggcgc acggggcttc ttcccatctt catgtgtccg cgagctctgc
     1741 ctctcctcac agagccggca gtggcactcc cagagcgccc tgtttcagat tccggaatat
     1801 tccatgggac aagcccgggc cctaatgggg ctttctgctc agctggatga agagctggac
     1861 ttcagggaag gggatgtgat caccattatt ggagttcctg aaccaggctg gtttgaaggg
     1921 gagttagagg gccgaagagg catttttcca gaaggttttg tagagctgtt ggggcccctg
     1981 aggactgtgg atgagtcagt aagttctgga aatcaagatg actgcattgt taatggtgaa
     2041 gtagataccc ccgtaggaga agaagagata gggccggatg aggatgagga ggagccaggg
     2101 acctatgggg tcgccctgta cagattccaa gccctggagc caaatgagct ggatttcgag
     2161 gtcggggata aaatccgaat tctggcgacc ttggaagatg gctggctgga aggatccctg
     2221 aagggcagga caggcatctt tccttaccgg tttgtgaaat tatgtcctga cacacgggtg
     2281 gaggaaacca tggctctgcc ccaggaaggc agccttgcca ggatcccgga aacttctttg
     2341 gattgtttgg agaacacctt aggagtagag gaacaaagac atgaaaccag tgaccatgag
     2401 gccgaggagc ctgactgcat tatttctgaa gcacccactt ctcccctcgg tcatctgact
     2461 tcagagtatg acacagacag aaactcttat caggacgagg acaccgcagg agggcccccg
     2521 agaagcccag gcgtggagtg ggaaatgcct cttgccacag actctcccac atctgaccct
     2581 acagaagtag tcaatggtat ttcctcccaa cctcaggtcc cttttcatcc caacttgcag
     2641 aaaagccagt attattctac agtgggagga agccacccgc actcagaaca gtaccccgac
     2701 cttcttcccc tagaagcaag gactagagac tatgccagcc tacctcccaa aagaatgtat
     2761 tcccagctaa aaactcttca gaagccagtg ctccctcttt acaggggctc ttctgtttca
     2821 gcttcaaggg tagtcaaacc cagacaatca agtcctcagc tccacaacct agcaagttat
     2881 actaaaaagc accacacgtc cagtgtttac tccatctcag agagattgga gatgaagcct
     2941 ggtccgcaag cccaagggct tgttatggaa gcagcaacac attcacaggg agacggcagc
     3001 actgacctgg actcgaagct gacacaacag ctgatcgagt ttgagaagag cttggcaggg
     3061 cccggcacag agccagataa aattttacgc cacttttcaa tcatggactt taactctgag
     3121 aaggatattg tccgaggttc ctcaaagtta atcaccgagc aggagctgcc ggaaaggaga
     3181 aaggccctaa ggccaccgcc acctcgtccc tgtactccgg tatccacttc tccccatttg
     3241 ctggttgacc agaacctaaa acctgcacca cccttggtgg tgcgaccctc tcgcccagct
     3301 cccctgcctc cctcagcaca gcagagaacg aatgcggtat cccccaagct cctatctcga
     3361 caccgtccta cctgtgagac cttagaaaag gagggccctg gtcatatggg aaggagtctg
     3421 gaccagacct ccccatgccc cttagtgctg gtgaggattg aggaaatgga gcgggacttg
     3481 gatatgtaca gtagagctca agaagagcta aacctcatgc tggaggagaa gcaggatgaa
     3541 tcatcaagag cagagaccct cgaggatctc aagttctgtg aaagtaacat tgaaagtttg
     3601 aatatggaac tccagcaact aagagaaatg acgctcctct cctcccagtc ttcatcactg
     3661 gtggcccctt ctgggtctgt gtctgccgaa aatccagagc agaggatgct ggagaagaga
     3721 gccaaggtca tagaagaact tcttcagaca gaaagagact acattcggga tctggaaatg
     3781 tgtattgagc ggatcatggt acccatgcag caggcacagg taccaaacat tgattttgag
     3841 ggactttttg gaaatatgca gatggtgatt aaggtctcga agcaattatt ggctgctctg
     3901 gaaatcagcg atgctgtagg acctgtgttt cttggtcacc gggatgagct tgagggaaca
     3961 tacaagattt actgccagaa tcatgatgag gccattgcgc tgcttgaaat ctacgagaag
     4021 gatgagaaga tccagaagca tcttcaggac tccttggcag atctgaagag cctatacaac
     4081 gaatggggat gcacaaatta tattaacctg ggctccttcc tcatcaaacc agtacagaga
     4141 gtaatgcgtt acccgctgtt gctaatggag ttgctgaatt ccaccccaga atcccaccca
     4201 gataaagtgc ctttaaccaa tgcagtcctt gcggtcaagg aaatcaacgt taacattaat
     4261 gaatataaac ggcgaaagga cctggtcctc aagtaccgta agggtgatga agatagcctt
     4321 atggagaaaa tttccaaact gaacatccac tccatcatca agaaatccaa ccgagttagc
     4381 agtcacctga agcatctcac tggctttgct cctcagataa aagatgaagt atttgaagaa
     4441 acagaaaaaa acttccgaat gcaagaaaga ttgattaagt cttttatccg agacctgtct
     4501 ctctacctcc agcacatccg ggagtccgca tgtgtgaaag tggtggctgc tgtgagcatg
     4561 tgggatgtgt gcatggagag aggacaccgg gacctggagc agtttgagag ggtgcatcgc
     4621 tacatcagtg accagctctt cacaaacttt aaggagagga cagagcggct tgtcatctcc
     4681 cccttaaatc agttactgag catgtttaca gggccccata agctggtaca gaaacgcttt
     4741 gacaagctcc tggacttcta taactgtaca gaacgggcag aaaagctaaa ggacaagaag
     4801 accctggagg agctgcagtc ggcccggaac aactatgagg ccctgaatgc acagctgctg
     4861 gatgagctgc ccaagttcca ccagtacgcc cagggcctct tcaccaactg tgtccacggc
     4921 tatgctgaag cccactgtga ctttgtgcac caggctctgg agcaattaaa gccactgctt
     4981 tcgttactca aagtggctgg cagagaggga aaccttatcg ccatcttcca cgaagagcac
     5041 agcagagttc tgcagcaact ccaggttttt accttcttcc cggagtctct tccagctacc
     5101 aagaaaccgt ttgagaggaa aaccattgac cgccagtctg ctcgaaagcc actcctgggc
     5161 ctgccaagtt acatgctaca gtcagaagaa ctccgggcct ccctcctggc caggtatccc
     5221 cctgaaaaac tcttccaggc agaacggaac ttcaatgctg cccaagactt ggatgtctca
     5281 cttttggaag gtgacctggt gggtgtgatt aagaaaaaag accccatggg cagccagaac
     5341 cgctggctga ttgacaatgg agtcaccaaa ggcttcgtgt acagctcttt cctaaagccc
     5401 tacaatcctc gccgcagcca ctccgatgcc tccgtgggta gccactcctc cacagagtct
     5461 gagcacggca gctcctcccc caggttccca cgccagaaca gcggcagcac cctgaccttc
     5521 aaccccagca gcatggctgt atcctttacc tcggggtctt gccagaagca gcctcaagat
     5581 gcatctcctc cgccaaaaga atgtgaccaa ggaactctca gtgcatccct aaatccgagt
     5641 aattcagaga gtagtccttc cagatgccct tcagacccag actccacctc ccagccaagg
     5701 tcaggggact ctgcagatgt agctagagat gtaaagcagc ccactgccac gccgaggagc
     5761 taccggaact tcaggcatcc agaaatagtt ggctactccg taccaggacg aaatgggcaa
     5821 agtcaagacc tcgtcaaagg atgtgcaaga acagcccagg ctccggaaga cagaagtaca
     5881 gagccagatg gcagtgaggc agaaggcaac caggtctatt ttgctgtcta caccttcaag
     5941 gcacgaaacc caaatgagct gagcgtgtca gccaatcaga aactcaagat cctcgagttt
     6001 aaagatgtta caggaaatac agagtggtgg ttagctgagg ttaacgggaa gaagggctac
     6061 gttccctcca attatatccg caaaaccgag tacacctaat ctagataact gatcataatc
     6121 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     6181 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     6241 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6301 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     6361 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     6421 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     6481 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     6541 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     6601 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6661 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6721 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6781 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6841 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6901 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     6961 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     7021 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     7081 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     7141 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     7201 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     7261 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     7321 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     7381 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     7441 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     7501 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     7561 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     7621 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     7681 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     7741 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     7801 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     7861 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     7921 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     7981 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     8041 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     8101 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     8161 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     8221 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     8281 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     8341 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     8401 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     8461 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     8521 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     8581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     9001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     9061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     9121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     9181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     9241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     9301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     9361 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     9421 gccatgcat
//
