LOCUS       sequence-346368-        5674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively active, nucelar localized RhoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5674)
  AUTHORS   Mahlandt EK, Arts JJG, van der Meer WJ, van der Linden FH, Tol S, 
            van Buul JD, Gadella TWJ, Goedhart J
  TITLE     Visualizing endogenous Rho activity with an improved 
            localization-based, genetically encoded biosensor.
  JOURNAL   J Cell Sci. 2021 Aug 6. pii: 271216. doi: 10.1242/jcs.258823.
  PUBMED    34357388
REFERENCE   2  (bases 1 to 5674)
  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.
            2021 Aug 6. pii: 271216. doi: 10.1242/jcs.258823."
COMMENT     Sequence Label: H2A-mTurquoise-RHOA-G14V-deltaCAAX
FEATURES             Location/Qualifiers
     source          1..5674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      413..422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(419..421,422..424,425..1135)
                     /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(464..485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(725..744)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1072..1093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1833..1954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1870..1889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1924..1943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1961..2416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2310..2329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2443..2547
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2549..2906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2549..2740
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2572..2592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2757..2892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2941..3735
                     /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             2941..3735
                     /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(2995..3014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3605..3624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3923..3942)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3967..4014
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4343..4931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4832..4851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5106..5409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5410..5613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5564..5584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aattctgatg tcgggacgcg gcaagcaggg aggcaaagct cgcgccaaag ccaagacccg
       61 ctcttctcgt gccggtctcc agttccccgt gggccgagtg caccgactgc tccgcaaggg
      121 caactatgct gagcgggtcg gggccggcgc gccggtgtac ctggcggcgg tgctggagta
      181 cctgactgcc gagatcctgg agctggcggg caacgccgcc cgcgacaaca agaagacccg
      241 cattatcccg cgccacttgc agctggccat ccgcaacgac gaggagctca acaagctgct
      301 gggcaaagta accatcgctc agggtggtgt cctgcccaac atccaggctg tgctactgcc
      361 caagaagacc gagagtcacc acaaggccaa aggcaaacgg gatccaccgg tcgccaccat
      421 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      481 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      541 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      601 cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
      661 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      721 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      781 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      841 gctggagtac aactacatca gcgacaacgt ctatatcacc gccgacaagc agaagaacgg
      901 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
      961 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1021 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1081 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccat
     1141 ggctgccatc cggaagaaac tggtgattgt tggtgatgta gcctgtggaa agacatgctt
     1201 gctcatagtc ttcagcaagg accagttccc agaggtgtat gtgcccacag tgtttgagaa
     1261 ctatgtggca gatatcgagg tggatggaaa gcaggtagag ttggctttgt gggacacagc
     1321 tgggcaggaa gattatgatc gcctgaggcc cctctcctac ccagataccg atgttatact
     1381 gatgtgtttt tccatcgaca gccctgatag tttagaaaac atcccagaaa agtggacccc
     1441 agaagtcaag catttctgtc ccaacgtgcc catcatcctg gttgggaata agaaggatct
     1501 tcggaatgat gagcacacaa ggcgggagct agccaagatg aagcaggagc cggtgaaacc
     1561 tgaagaaggc agagatatgg caaacaggat tggcgctttt gggtacatgg agtgttcagc
     1621 aaagaccaaa gatggagtga gagaggtttt tgaaatggct acgagagctg ctctgcaagc
     1681 tagacgtggg aagaaaaaat ctgggagcta agcggccgcg actctagatc ataatcagcc
     1741 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1801 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1861 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1921 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     1981 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2041 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2101 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2161 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2221 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2281 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2341 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2401 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2461 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2521 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     2581 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2641 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2761 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2821 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2881 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     2941 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3001 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3061 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3121 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3181 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3241 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3301 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3361 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3421 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     3481 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3541 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3601 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3661 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3721 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3781 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3841 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3901 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3961 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4021 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4081 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4141 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4201 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4261 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4321 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4381 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4441 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4501 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4561 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4621 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4681 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4741 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4801 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4861 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4921 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4981 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5041 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5101 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5161 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5221 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     5281 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     5341 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     5401 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     5461 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     5521 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     5581 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
     5641 cgctaccgga ctcagatctc gagctcaagc ttcg
//
