LOCUS       sequence-346392-        4988 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 4988)
  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 4988)
  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: mTurquoise2-3xrGBD
FEATURES             Location/Qualifiers
     source          1..4988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(159..161,162..164,165..872)
                     /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(204..225)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(465..484)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     812..833
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1776..1897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1813..1832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1867..1886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1904..2359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2041..2062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2253..2272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2386..2490
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2492..2849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2492..2683
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2515..2535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2700..2835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2884..3678
                     /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             2884..3678
                     /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(2938..2957)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3548..3567
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3866..3885)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3910..3957
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4286..4874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4775..4794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tgggaggtct atataagcag agctggttta
      121 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      181 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      241 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      301 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgtcctggg
      361 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      421 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      481 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      541 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacttta
      601 gcgacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
      661 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
      721 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
      781 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
      841 ccgggatcac tctcggcatg gacgagctgt acatccccct cgagtccatc ctggaggacc
      901 tcaatatgct ctacatccgg cagatggcac tcagcctgga ggacacagag ctgcagagga
      961 aactagatca tgagatccgg atgagggatg gggcctgcaa gctgctggca gcctgctccc
     1021 agcgagagca ggctctggaa gccaccaaga gcctgctggt gtgcaacagc cgtattctca
     1081 gctacatggg tgagctgcag cggcgaaagg aggcccaggt gctggagaag acaggcaggg
     1141 tcgagtccat cctggaggac ctcaatatgc tctacatccg gcagatggca ctcagcctgg
     1201 aggacacaga gctgcagagg aaactagatc atgagatccg gatgagggat ggggcctgca
     1261 agctgctggc agcctgctcc cagcgagagc aggctctgga agccaccaag agcctgctgg
     1321 tgtgcaacag ccgtattctc agctacatgg gtgagctgca gcggcgaaag gaggcccagg
     1381 tgctggagaa gacaggcagg cgtcgagtta tcctggagga cctcaatatg ctctacatcc
     1441 ggcagatggc actcagcctg gaggacacag agctgcagag gaaactagat catgagatcc
     1501 ggatgaggga tggggcctgc aagctgctgg cagcctgctc ccagcgagag caggctctgg
     1561 aagccaccaa gagcctgctg gtgtgcaaca gccgtattct cagctacatg ggtgagctgc
     1621 agcggcgaaa ggaggcccag gtgctggaga agacaggctc tagataactg atcataatca
     1681 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1741 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     1921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2341 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2401 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2461 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2521 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2581 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2641 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2701 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2761 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2821 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     2881 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2941 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3001 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3061 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3121 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3181 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3241 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3301 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3361 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3421 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3481 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3541 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3601 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3661 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3721 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3781 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3841 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3901 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     3961 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4021 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4081 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4141 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4201 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4921 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4981 ccatgcat
//
