LOCUS       sequence-346393-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively active, nuclear localized Cdc42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  AUTHORS   Mahlandt EK, Arts JJG, van der Meer WJ, van der Linden FH, Tol S, 
            van Buul JD, Gadella TWJ, Goedhart J
  TITLE     Visualizing endogenous Rho activity with an improved 
            localization-based, genetically encoded biosensor.
  JOURNAL   J Cell Sci. 2021 Aug 6. pii: 271216. doi: 10.1242/jcs.258823.
  PUBMED    34357388
REFERENCE   2  (bases 1 to 5668)
  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: "J Cell Sci.
            2021 Aug 6. pii: 271216. doi: 10.1242/jcs.258823."
COMMENT     Sequence Label: H2A-mTurquoise-CDC42-G12V-deltaCAAX
FEATURES             Location/Qualifiers
     source          1..5668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      413..422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(419..421,422..424,425..1135)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(464..485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(725..744)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1072..1093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1827..1948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1864..1883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1918..1937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1955..2410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2092..2113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2304..2323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2437..2541
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2543..2900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2543..2734
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2566..2586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2751..2886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2935..3729
                     /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             2935..3729
                     /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(2989..3008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3599..3618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3917..3936)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3961..4008
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4337..4925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4826..4845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5100..5403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5404..5607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5558..5578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aattctgatg tcgggacgcg gcaagcaggg aggcaaagct cgcgccaaag ccaagacccg
       61 ctcttctcgt gccggtctcc agttccccgt gggccgagtg caccgactgc tccgcaaggg
      121 caactatgct gagcgggtcg gggccggcgc gccggtgtac ctggcggcgg tgctggagta
      181 cctgactgcc gagatcctgg agctggcggg caacgccgcc cgcgacaaca agaagacccg
      241 cattatcccg cgccacttgc agctggccat ccgcaacgac gaggagctca acaagctgct
      301 gggcaaagta accatcgctc agggtggtgt cctgcccaac atccaggctg tgctactgcc
      361 caagaagacc gagagtcacc acaaggccaa aggcaaacgg gatccaccgg tcgccaccat
      421 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      481 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      541 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      601 cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
      661 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      721 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      781 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      841 gctggagtac aactacatca gcgacaacgt ctatatcacc gccgacaagc agaagaacgg
      901 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
      961 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1021 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1081 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccat
     1141 gcagacaatt aagtgtgttg ttgtgggcga tgttgctgtt ggtaaaacat gtctcctgat
     1201 atcctacaca acaaacaaat ttccatcgga atatgtaccg actgtttttg acaactatgc
     1261 agtcacagtt atgattggtg gagaaccata tactcttgga ctttttgata ctgcagggca
     1321 agaggattat gacagattac gaccgctgag ttatccacaa acagatgtat ttctagtctg
     1381 tttttcagtg gtctctccat cttcatttga aaacgtgaaa gaaaagtggg tgcctgagat
     1441 aactcaccac tgtccaaaga ctcctttctt gcttgttggg actcaaattg atctcagaga
     1501 tgacccctct actattgaga aacttgccaa gaacaaacag aagcctatca ctccagagac
     1561 tgctgaaaag ctggcccgtg acctgaaggc tgtcaagtat gtggagtgtt ctgcacttac
     1621 acagagaggt ctgaagaatg tgtttgatga ggctatccta gctgccctcg agcctccgga
     1681 aactcaaccc aaaaggaaga gctaagcggc cgcgactcta gatcataatc agccatacca
     1741 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1801 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1861 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1921 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     1981 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2041 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2101 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2161 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2221 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2281 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2341 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2401 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2461 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2521 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2581 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2641 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2761 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2821 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2881 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2941 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3001 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3061 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3121 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3181 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3241 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3301 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3361 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3421 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3481 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3541 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3601 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3661 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3721 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3781 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3841 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3901 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3961 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4021 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4081 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4141 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4201 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4261 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4321 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4381 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4441 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4501 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4561 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4621 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4681 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4741 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4801 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4861 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4921 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4981 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     5041 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5101 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5161 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5221 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5281 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5341 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5401 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5461 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5521 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5581 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     5641 cggactcaga tctcgagctc aagcttcg
//
