LOCUS       sequence-261567-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of active RhoA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4442)
  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 4442)
  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: mNeongreen-aGBD(anillin)
FEATURES             Location/Qualifiers
     source          1..4442
                     /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"
     polyA_signal    1230..1351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1267..1286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1321..1340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1358..1813)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1495..1516)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1707..1726
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1840..1944
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1946..2303
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1946..2137
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1969..1989)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2154..2289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2338..3132
                     /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             2338..3132
                     /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(2392..2411)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3002..3021
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3320..3339)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3364..3411
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3740..4328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4229..4248
                     /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 tacagcctcg agcaagttaa tatcaaacag aaaatgcagg
      901 aactcaataa cgaaataaat atgcaacaga cagtgatcta tcaagctagc caggctctta
      961 actgctgtgt tgatgaagaa catggaaaag ggtccctaga agaagctgaa gcagaaagac
     1021 ttcttctaat tgcaactggg aagagaacac ttttgattga tgaattgaat aaattgaaga
     1081 acgaaggacc tcagaggtaa ggatccaccg gatctagata actgatcata atcagccata
     1141 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     1201 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     1261 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1321 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     1381 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1441 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1501 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1561 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1621 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1681 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1741 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     1801 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1921 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     1981 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2041 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2101 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2161 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2221 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2281 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     2341 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2401 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2461 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     2521 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     2581 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     2641 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     2701 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     2761 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     2821 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     2881 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     2941 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3001 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3061 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3121 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3181 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3241 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3301 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3361 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     3421 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     3481 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     3541 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     3601 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3661 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3721 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3781 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3841 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3901 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3961 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4021 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4081 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4141 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4201 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4261 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4321 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4381 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     4441 at
//
