LOCUS       sequence-346254-        3832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of active Rho.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3832)
  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 3832)
  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: pEntra1a-mTurquoise2-3xrGBD
FEATURES             Location/Qualifiers
     source          1..3832
                     /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      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(506..508,509..511,512..1219)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(551..572)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(812..831)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1159..1180
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2061..2160)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2272..2291)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2283..3092
                     /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(2350..2369)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3182..3770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3671..3690
                     /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 accccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      541 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      601 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      661 caagctgccc gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt
      721 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      781 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      841 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      901 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta
      961 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1021 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1081 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     1141 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1201 cggcatggac gagctgtaca tccccctcga gtccatcctg gaggacctca atatgctcta
     1261 catccggcag atggcactca gcctggagga cacagagctg cagaggaaac tagatcatga
     1321 gatccggatg agggatgggg cctgcaagct gctggcagcc tgctcccagc gagagcaggc
     1381 tctggaagcc accaagagcc tgctggtgtg caacagccgt attctcagct acatgggtga
     1441 gctgcagcgg cgaaaggagg cccaggtgct ggagaagaca ggcagggtcg agtccatcct
     1501 ggaggacctc aatatgctct acatccggca gatggcactc agcctggagg acacagagct
     1561 gcagaggaaa ctagatcatg agatccggat gagggatggg gcctgcaagc tgctggcagc
     1621 ctgctcccag cgagagcagg ctctggaagc caccaagagc ctgctggtgt gcaacagccg
     1681 tattctcagc tacatgggtg agctgcagcg gcgaaaggag gcccaggtgc tggagaagac
     1741 aggcaggcgt cgagttatcc tggaggacct caatatgctc tacatccggc agatggcact
     1801 cagcctggag gacacagagc tgcagaggaa actagatcat gagatccgga tgagggatgg
     1861 ggcctgcaag ctgctggcag cctgctccca gcgagagcag gctctggaag ccaccaagag
     1921 cctgctggtg tgcaacagcc gtattctcag ctacatgggt gagctgcagc ggcgaaagga
     1981 ggcccaggtg ctggagaaga caggctctag ataactgatc ataatcagcc ataccacgcg
     2041 gccgcactcg agatatctag acccagcttt cttgtacaaa gttggcatta taagaaagca
     2101 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2161 ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2221 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2281 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2341 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2401 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2461 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2521 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2581 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2641 atgcgctggc agtgtccctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2701 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2761 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2821 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2881 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2941 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3001 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3061 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacatt
     3121 attcagattg ggccccgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3301 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3361 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3421 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3481 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3541 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3601 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3661 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3721 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3781 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
