LOCUS       sequence-260228-        4718 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 4718)
  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 4718)
  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-2xrGBD
FEATURES             Location/Qualifiers
     source          1..4718
                     /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    1506..1627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1543..1562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1597..1616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1634..2089)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1771..1792)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1983..2002
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2116..2220
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2222..2579
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2222..2413
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2245..2265)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2430..2565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2614..3408
                     /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             2614..3408
                     /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(2668..2687)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3278..3297
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3596..3615)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3640..3687
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4016..4604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4505..4524
                     /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 tacatccccc tcgagtccat cctggaggac ctcaatatgc
      901 tctacatccg gcagatggca ctcagcctgg aggacacaga gctgcagagg aaactagatc
      961 atgagatccg gatgagggat ggggcctgca agctgctggc agcctgctcc cagcgagagc
     1021 aggctctgga agccaccaag agcctgctgg tgtgcaacag ccgtattctc agctacatgg
     1081 gtgagctgca gcggcgaaag gaggcccagg tgctggagaa gacaggcagg cgtcgagtta
     1141 tcctggagga cctcaatatg ctctacatcc ggcagatggc actcagcctg gaggacacag
     1201 agctgcagag gaaactagat catgagatcc ggatgaggga tggggcctgc aagctgctgg
     1261 cagcctgctc ccagcgagag caggctctgg aagccaccaa gagcctgctg gtgtgcaaca
     1321 gccgtattct cagctacatg ggtgagctgc agcggcgaaa ggaggcccag gtgctggaga
     1381 agacaggctc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     1441 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1501 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1561 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1621 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     1681 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1741 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1801 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     1861 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     1921 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     1981 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2041 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2101 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2161 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2221 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2281 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2341 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2401 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2461 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2521 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     2581 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2641 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2701 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2761 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     2821 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2881 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2941 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3001 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3061 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3121 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3181 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3241 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3301 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3361 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3421 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3481 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3541 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     3601 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     3661 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     3721 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     3781 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     3841 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     3901 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3961 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4021 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4081 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4141 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4201 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4261 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4321 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4381 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4441 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4501 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4561 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4621 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4681 cccctgattc tgtggataac cgtattaccg ccatgcat
//
