LOCUS       sequence-346331-        5688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutivley active, mitochondrial localized Cdc42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  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 5688)
  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: TOMM20-mTurquoise-Cdc42-G12V-DeltaCaaX
FEATURES             Location/Qualifiers
     source          1..5688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      363..372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      828..837
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(834..836,837..839,840..1550)
                     /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(879..900)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1140..1159)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1487..1508
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2242..2363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2279..2298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2333..2352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2370..2825)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2507..2528)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2719..2738
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2852..2956
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2958..3315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2958..3149
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2981..3001)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3166..3301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3350..4144
                     /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             3350..4144
                     /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(3404..3423)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4014..4033
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4332..4351)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4376..4423
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4752..5340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5241..5260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5515..5688,1..130)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgccaccat ggtgggtcgg aacagcgcca tcgccgccgg tgtatgcggg gcccttttca
      421 ttgggtactg catctacttc gaccgcaaaa gacgaagtga ccccaacttc aagaacaggc
      481 ttcgagaacg aagaaagaaa cagaagcttg ccaaggagag agctgggctt tccaagttac
      541 ctgaccttaa agatgctgaa gctgttcaga agttcttcct tgaagaaata cagcttggtg
      601 aagagttact agctcaaggt gaatatgaga agggcgtaga ccatctgaca aatgcaattg
      661 ctgtgtgtgg acagccacag cagttactgc aggtcttaca gcaaactctt ccaccaccag
      721 tgttccagat gcttctgact aagctcccaa caattagtca gagaattgta agtgctcaga
      781 gcttggctga agatgatgtg gaaggcggta gcggggatcc accggtcgcc accatggtga
      841 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      901 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      961 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1021 ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1081 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1141 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1201 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1261 agtacaacta catcagcgac aacgtctata tcaccgccga caagcagaag aacggcatca
     1321 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1381 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1441 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1501 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccatgcaga
     1561 caattaagtg tgttgttgtg ggcgatgttg ctgttggtaa aacatgtctc ctgatatcct
     1621 acacaacaaa caaatttcca tcggaatatg taccgactgt ttttgacaac tatgcagtca
     1681 cagttatgat tggtggagaa ccatatactc ttggactttt tgatactgca gggcaagagg
     1741 attatgacag attacgaccg ctgagttatc cacaaacaga tgtatttcta gtctgttttt
     1801 cagtggtctc tccatcttca tttgaaaacg tgaaagaaaa gtgggtgcct gagataactc
     1861 accactgtcc aaagactcct ttcttgcttg ttgggactca aattgatctc agagatgacc
     1921 cctctactat tgagaaactt gccaagaaca aacagaagcc tatcactcca gagactgctg
     1981 aaaagctggc ccgtgacctg aaggctgtca agtatgtgga gtgttctgca cttacacaga
     2041 gaggtctgaa gaatgtgttt gatgaggcta tcctagctgc cctcgagcct ccggaaactc
     2101 aacccaaaag gaagagctaa gcggccgcga ctctagatca taatcagcca taccacattt
     2161 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2221 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2341 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     2401 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2461 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2521 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2581 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2641 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2701 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2761 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2821 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2881 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2941 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3061 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3181 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3241 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3301 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3361 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3421 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3481 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3541 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3601 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3661 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3721 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3781 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3841 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3901 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3961 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4021 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4081 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4141 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4201 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4261 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4321 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4381 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4441 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4501 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4561 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4621 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5401 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     5461 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5521 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5581 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5641 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatca
//
