LOCUS       sequence-346324-        5691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively active, mitochondrial localized Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5691)
  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 5691)
  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-Rac1-G12V-deltaCaaX
FEATURES             Location/Qualifiers
     source          1..5691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        133..336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     287..307
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      365..374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      830..839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(836..838,839..841,842..1552)
                     /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(881..902)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1142..1161)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1489..1510
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2247..2368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2284..2303)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2338..2357
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2375..2830)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2512..2533)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2724..2743
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2857..2961
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2963..3320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2963..3154
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2986..3006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3171..3306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3355..4149
                     /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             3355..4149
                     /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(3409..3428)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4019..4038
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4337..4356)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4381..4428
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4757..5345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5246..5265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5520..5691,1..132)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
       61 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      121 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      181 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      241 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      301 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      361 tagcgccacc atggtgggtc ggaacagcgc catcgccgcc ggtgtatgcg gggccctttt
      421 cattgggtac tgcatctact tcgaccgcaa aagacgaagt gaccccaact tcaagaacag
      481 gcttcgagaa cgaagaaaga aacagaagct tgccaaggag agagctgggc tttccaagtt
      541 acctgacctt aaagatgctg aagctgttca gaagttcttc cttgaagaaa tacagcttgg
      601 tgaagagtta ctagctcaag gtgaatatga gaagggcgta gaccatctga caaatgcaat
      661 tgctgtgtgt ggacagccac agcagttact gcaggtctta cagcaaactc ttccaccacc
      721 agtgttccag atgcttctga ctaagctccc aacaattagt cagagaattg taagtgctca
      781 gagcttggct gaagatgatg tggaaggcgg tagcggggat ccaccggtcg ccaccatggt
      841 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      901 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      961 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1021 gaccaccctg tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1081 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1141 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1201 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1261 ggagtacaac tacatcagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1321 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     1381 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1441 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1501 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccatgca
     1561 ggccatcaag tgtgtggtgg tgggagacgt agctgtaggt aaaacttgcc tactgatcag
     1621 ttacacaacc aatgcatttc ctggagaata tatccctact gtctttgaca attattctgc
     1681 caatgttatg gtagatggaa aaccggtgaa tctgggctta tgggatacag ctggacaaga
     1741 agattatgac agattacgcc ccctatccta tccgcaaaca gatgtgttct taatttgctt
     1801 ttcccttgtg agtcctgcat catttgaaaa tgtccgtgca aagtggtatc ctgaggtgcg
     1861 gcaccactgt cccaacactc ccatcatcct agtgggaact aaacttgatc ttagggatga
     1921 taaagacacg atcgagaaac tgaaggagaa gaagctgact cccatcacct atccgcaggg
     1981 tctagccatg gctaaggaga ttggtgctgt aaaatacctg gagtgctcgg cgctcacaca
     2041 gcgaggcctc aagacagtgt ttgacgaagc gatccgagca gtcctctgcc cgcctcccgt
     2101 gaagaagagg aagagaaaaa gctaagcggc cgcgactcta gatcataatc agccatacca
     2161 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2221 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2281 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2341 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2401 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2461 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2521 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2581 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2641 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2701 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2761 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2821 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2881 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2941 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3001 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3181 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3241 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3301 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3361 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3421 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3481 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3541 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3601 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3661 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3721 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3781 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3841 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3901 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3961 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4021 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4081 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4141 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4201 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4261 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4321 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4381 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4441 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4501 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4561 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4621 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4681 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4741 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4801 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4861 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4921 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4981 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5041 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5101 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5161 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5221 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5281 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5341 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5401 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     5461 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5521 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5581 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5641 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta t
//
