LOCUS       sequence-261585-        7341 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 7341)
  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 7341)
  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-SGEF
FEATURES             Location/Qualifiers
     source          1..7341
                     /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    4129..4250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4166..4185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4220..4239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4257..4712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4394..4415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4606..4625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4739..4843
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4845..5202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4845..5036
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4868..4888)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5053..5188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5237..6031
                     /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             5237..6031
                     /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(5291..5310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5901..5920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6219..6238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6263..6310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6639..7227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7128..7147
                     /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 tcaagcttcc gaattcatgg acggcgagag cgaggtggat
     1381 ttttctagca acagcataac ccctttgtgg cggaggcggt cgattcctca gccccaccag
     1441 cttctgggcc ggagcaagcc gaggccccag tcctaccaga gccccaacgg gttactaatt
     1501 acggatttcc cggtggagga cggagggacg ctctccgcag cgcagattcc cgcccaggtg
     1561 cccaccgcct cggacagcag gacggtacat aggagccccc tgcttctggg cgcccagcgg
     1621 agagcggtgg ccaatggtgg gacggcatcc ccggagtaca gggctgcctc tcctcgactt
     1681 cgacggccca agtcacccaa gctccccaaa gcggtgcctg gcggctcccc gaaatcccca
     1741 gcaaatggcg cggtgacctt gcctgcgccg ccgccgccgc cggttctgcg ccccccgcgg
     1801 actcctaacg cgcccgcccc ctgcaccccc gaggaggacc ttactgggtt gactgccagc
     1861 ccggtgcctt cgcccactgc aaatggcctt gccgctaata acgactctcc tgggtcaggt
     1921 tcgcagtccg gccggaaggc aaaggacccc gaacgggggc tctttcctgg gccccagaaa
     1981 agttcttcgg aacaaaaact ccccctccaa aggctgccct cccaggagaa cgagctcctc
     2041 gagaatcctt ccgtggtttt gagtacaaac agccccgccg ccctcaaagt ggggaagcag
     2101 cagatcattc cgaagagtct ggcctcggaa attaaaataa gtaaatccaa caatcaaaat
     2161 gtggagcccc acaagagact cctcaaggtg cgcagcatgg tggagggcct aggaggaccc
     2221 ctgggtcacg caggggagga gagtgaggtc gataacgacg tggatagccc agggtctctg
     2281 cggagaggct tgcggtccac gtcttatcgc agggcagtgg tcagtggctt tgattttgac
     2341 agtcctacca gctcgaagaa gaagaacaga atgtcccagc ctgttctgaa agtggtgatg
     2401 gaagacaagg agaagttttc cagtctggga aggataaaga aaaaaatgct gaaaggacaa
     2461 ggaacatttg atggggaaga aaatgctgtc ctgtatcaaa actacaagga aaaggccctt
     2521 gacattgatt ctgatgaaga gtcagagccc aaagaacaga agtcagatga aaaaattgtg
     2581 attcaccata agccattgag atccacatgg agccaactct ctgcggtgaa aagaaaggga
     2641 ttatctcaga cagtaagcca ggaggaaaga aagagacaag aggctatctt tgaagtcata
     2701 tcctctgaac attcatattt actcagcttg gagatcttga tacgaatgtt taaaaattct
     2761 aaagaactga gtgatacaat gactaaaacc gagaggcacc atcttttctc caatattaca
     2821 gatgtctgtg aggcaagcaa aaagttcttt atagagttgg aagcaagaca tcagaataat
     2881 atcttcatag atgacataag tgacattgtg gaaaaacaca cagcatccac atttgaccca
     2941 tatgtgaaat actgcacaaa tgaagtctac caacaacgaa cactacaaaa attgttagct
     3001 accaatccat cctttaagga agtattgtca aggattgagt cccatgaaga ctgtaggaac
     3061 ttacccatga tctcttttct cattctcccc atgcagaggg tgacccgcct tcccctgctg
     3121 atggatacta tctgtcaaaa aacacctaag gactctccga agtatgaagt ctgcaaaaga
     3181 gccttgaagg aagttagcaa gttggttcga ctatgcaatg agggcgcccg gaagatggaa
     3241 aggactgaga tgatgtacac aattaactcc cagctggaat ttaaaattaa gccttttcct
     3301 ttagtctcct cttcccggtg gttggtaaaa agaggtgaat tgacagccta tgttgaagac
     3361 actgtgcttt tctcaagaag gacatccaaa cagcaagtct acttctttct ctttaacgat
     3421 gtgctcatta tcaccaagaa gaagagtgaa gaaagttaca acgtcaatga ttattcctta
     3481 agagatcagc tattggtgga atcttgtgac aatgaagagc ttaattcttc tccagggaag
     3541 aacagctcca caatgctcta ttcaagacag agctctgcca gtcacctctt tactctgaca
     3601 gtccttagta accacgcgaa tgagaaagtg gagatgctac taggagctga gacgcagagc
     3661 gagcgagccc gctggataac tgccctggga cacagcagcg ggaagccgcc tgcagaccga
     3721 acctcactga cccaggtgga aatcgttagg tcatttactg ctaagcagcc agatgaactc
     3781 tccctgcagg tggctgacgt cgtcctcatc tatcaacgtg tcagcgatgg ctggtatgag
     3841 ggggaacgac tacgagatgg agaaagaggc tggtttccta tggaatgtgc caaggagata
     3901 acatgtcaag ctacaattga taagaatgtg gagagaatgg gacgcttgct aggactggag
     3961 accaacgtgt agggatccgt cgacggtacc gcgggcccgg gatccaccgg atctagataa
     4021 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4081 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4141 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4201 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     4261 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4321 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4381 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4441 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4501 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4561 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4621 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4681 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4741 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4801 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4861 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4921 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4981 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5041 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5101 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5161 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     5221 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5281 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5341 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5401 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5461 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5521 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5581 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5641 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5701 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5761 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5821 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5881 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5941 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6001 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6061 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6121 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6181 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     6241 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     6301 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6361 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6421 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6481 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6781 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7321 aaccgtatta ccgccatgca t
//
