LOCUS       sequence-346312-        6425 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 6425)
  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 6425)
  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: 3xmNeonGreen-3xrGBD
FEATURES             Location/Qualifiers
     source          1..6425
                     /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             159..863
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      414..423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             882..1586
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1137..1146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1605..2309
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1860..1869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3213..3334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3250..3269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3304..3323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3341..3796)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3478..3499)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3690..3709
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3823..3927
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3929..4286
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3929..4120
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3952..3972)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4137..4272
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4321..5115
                     /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             4321..5115
                     /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(4375..4394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4985..5004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5303..5322)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5347..5394
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5723..6311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6212..6231
                     /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 ggcgaggagg
      181 ataacatggc ctctctccca gcgacacatg agttacacat ctttggctcc atcaacggtg
      241 tggactttga catggtgggt cagggcaccg gcaatccaaa tgatggttat gaggagttaa
      301 acctgaagtc caccaagggt gacctccagt tctccccctg gattctggtc cctcatatcg
      361 ggtatggctt ccatcagtac ctgccctacc ctgacgggat gtcgcctttc caggccgcca
      421 tggtagatgg ctccggctac caagtccatc gcacaatgca gtttgaagat ggtgcctccc
      481 ttactgttaa ctaccgctac acctacgagg gaagccacat caaaggagag gcccaggtga
      541 aggggactgg tttccctgct gacggtcctg tgatgaccaa ctcgctgacc gctgcggact
      601 ggtgcaggtc gaagaagact taccccaacg acaaaaccat catcagtacc tttaagtgga
      661 gttacaccac tggaaatggc aagcgctacc ggagcactgc gcggaccacc tacacctttg
      721 ccaagccaat ggcggctaac tatctgaaga accagccgat gtacgtgttc cgtaagacgg
      781 agctcaagca ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg
      841 tgatgggcat ggacgagctg taccagggtg gcagcggtgg catggtgagc aagggcgagg
      901 aggataacat ggcctctctc ccagcgacac atgagttaca catctttggc tccatcaacg
      961 gtgtggactt tgacatggtg ggtcagggca ccggcaatcc aaatgatggt tatgaggagt
     1021 taaacctgaa gtccaccaag ggtgacctcc agttctcccc ctggattctg gtccctcata
     1081 tcgggtatgg cttccatcag tacctgccct accctgacgg gatgtcgcct ttccaggccg
     1141 ccatggtaga tggctccggc taccaagtcc atcgcacaat gcagtttgaa gatggtgcct
     1201 cccttactgt taactaccgc tacacctacg agggaagcca catcaaagga gaggcccagg
     1261 tgaaggggac tggtttccct gctgacggtc ctgtgatgac caactcgctg accgctgcgg
     1321 actggtgcag gtcgaagaag acttacccca acgacaaaac catcatcagt acctttaagt
     1381 ggagttacac cactggaaat ggcaagcgct accggagcac tgcgcggacc acctacacct
     1441 ttgccaagcc aatggcggct aactatctga agaaccagcc gatgtacgtg ttccgtaaga
     1501 cggagctcaa gcactccaag accgagctca acttcaagga gtggcaaaag gcctttaccg
     1561 atgtgatggg catggacgag ctgtaccagg gtggcagcgg tggcatggtg agcaagggcg
     1621 aggaggataa catggcctct ctcccagcga cacatgagtt acacatcttt ggctccatca
     1681 acggtgtgga ctttgacatg gtgggtcagg gcaccggcaa tccaaatgat ggttatgagg
     1741 agttaaacct gaagtccacc aagggtgacc tccagttctc cccctggatt ctggtccctc
     1801 atatcgggta tggcttccat cagtacctgc cctaccctga cgggatgtcg cctttccagg
     1861 ccgccatggt agatggctcc ggctaccaag tccatcgcac aatgcagttt gaagatggtg
     1921 cctcccttac tgttaactac cgctacacct acgagggaag ccacatcaaa ggagaggccc
     1981 aggtgaaggg gactggtttc cctgctgacg gtcctgtgat gaccaactcg ctgaccgctg
     2041 cggactggtg caggtcgaag aagacttacc ccaacgacaa aaccatcatc agtaccttta
     2101 agtggagtta caccactgga aatggcaagc gctaccggag cactgcgcgg accacctaca
     2161 cctttgccaa gccaatggcg gctaactatc tgaagaacca gccgatgtac gtgttccgta
     2221 agacggagct caagcactcc aagaccgagc tcaacttcaa ggagtggcaa aaggccttta
     2281 ccgatgtgat gggcatggac gagctgtaca tccccctcga gtccatcctg gaggacctca
     2341 atatgctcta catccggcag atggcactca gcctggagga cacagagctg cagaggaaac
     2401 tagatcatga gatccggatg agggatgggg cctgcaagct gctggcagcc tgctcccagc
     2461 gagagcaggc tctggaagcc accaagagcc tgctggtgtg caacagccgt attctcagct
     2521 acatgggtga gctgcagcgg cgaaaggagg cccaggtgct ggagaagaca ggcagggtcg
     2581 agtccatcct ggaggacctc aatatgctct acatccggca gatggcactc agcctggagg
     2641 acacagagct gcagaggaaa ctagatcatg agatccggat gagggatggg gcctgcaagc
     2701 tgctggcagc ctgctcccag cgagagcagg ctctggaagc caccaagagc ctgctggtgt
     2761 gcaacagccg tattctcagc tacatgggtg agctgcagcg gcgaaaggag gcccaggtgc
     2821 tggagaagac aggcaggcgt cgagttatcc tggaggacct caatatgctc tacatccggc
     2881 agatggcact cagcctggag gacacagagc tgcagaggaa actagatcat gagatccgga
     2941 tgagggatgg ggcctgcaag ctgctggcag cctgctccca gcgagagcag gctctggaag
     3001 ccaccaagag cctgctggtg tgcaacagcc gtattctcag ctacatgggt gagctgcagc
     3061 ggcgaaagga ggcccaggtg ctggagaaga caggctctag ataactgatc ataatcagcc
     3121 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3181 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3241 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3301 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3361 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3421 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3481 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3541 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3601 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3661 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3721 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3781 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3901 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3961 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4141 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4201 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4261 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4321 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4381 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4441 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4501 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4561 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4621 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4681 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4741 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4801 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4861 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4921 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4981 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5041 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5101 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5161 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5221 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5281 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5341 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5401 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5461 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5521 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5581 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5641 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5701 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5761 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5821 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5881 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5941 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6001 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6061 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6121 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6181 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6241 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6301 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6361 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6421 tgcat
//
