LOCUS       sequence-346252-        3631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of active Cdc42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3631)
  AUTHORS   Mahlandt EK, Kreider-Letterman G, Chertkova AO, Garcia-Mata R, 
            Goedhart J
  TITLE     Cell-based optimization and characterization of genetically encoded 
            location-based biosensors for Cdc42 or Rac activity.
  JOURNAL   J Cell Sci. 2023 May 15;136(10):jcs260802. doi: 10.1242/jcs.260802. 
            Epub 2023 May 25.
  PUBMED    37226883
REFERENCE   2  (bases 1 to 3631)
  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."; date: "2023-05-15"; volume: "136(10):jcs260802. doi"; pages: 
            " 10.1242/jcs.260802. Epub 2023 May 25"
COMMENT     Sequence Label: pEntr1a-wGBD-mTurquoise2-wGBD
FEATURES             Location/Qualifiers
     source          1..3631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      801..810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(807..809,810..812,813..1523)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(852..873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1113..1132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1460..1481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1860..1959)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2071..2090)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2082..2891
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2149..2168)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2981..3569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3470..3489
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt acccgtcaga tccgctagca tgcctagccc agctgataag aaacgctcag
      541 ggaagaagaa gatcagcaaa gctgatattg gtgcacccag tggattcaag catgtcagcc
      601 acgtggggtg ggacccccag aatggatttg acgtgaacaa cctcgaccca gatctgcgga
      661 gtctgttctc cagggcagga atcagcgagg cccagctcac cgacgccgag acctctaaac
      721 ttatctacga cttcattgag gaccagggtg ggctggaggc tgtgcggcag gagatgaggc
      781 gccaggagtt ctcaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      841 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      901 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      961 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gtcctggggc gtgcagtgct
     1021 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1081 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1141 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1201 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctactttagc gacaacgtct
     1261 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1321 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1381 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     1441 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1501 tcggcatgga cgagctgtac aagtccggac ctagcccagc tgataagaaa cgctcaggga
     1561 agaagaagat cagcaaagct gatattggtg cacccagtgg attcaagcat gtcagccacg
     1621 tggggtggga cccccagaat ggatttgacg tgaacaacct cgacccagat ctgcggagtc
     1681 tgttctccag ggcaggaatc agcgaggccc agctcaccga cgccgagacc tctaaactta
     1741 tctacgactt cattgaggac cagggtgggc tggaggctgt gcggcaggag atgaggcgcc
     1801 aggagttcta gataactgat cataatcagc cataccgcgg ccgcactcga gatatctaga
     1861 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1921 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg cagctctggc
     1981 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     2041 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     2101 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2161 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     2221 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2281 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2341 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     2401 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgtccctgc
     2461 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2521 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2581 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     2641 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2701 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2761 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2821 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2881 agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg gccccgttcc
     2941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3121 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3601 tggccttttg ctggcctttt gctcacatgt t
//
