LOCUS       sequence-263526-        4430 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 4430)
  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 4430)
  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-pGBD(PKN1 codon optimized)
FEATURES             Location/Qualifiers
     source          1..4430
                     /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    1218..1339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1255..1274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1309..1328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1346..1801)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1483..1504)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1695..1714
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1828..1932
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1934..2291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1934..2125
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1957..1977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2142..2277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2326..3120
                     /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             2326..3120
                     /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(2380..2399)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2990..3009
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3308..3327)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3352..3399
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3728..4316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4217..4236
                     /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 agggggtaca gcagcagctg gagctggaaa
      901 gagaaaggtt aagaagagag atcagaaaag aattaaagct gaaggaagga gctgagaacc
      961 tgaggagagc caccacagat ttgggaagaa gcctgggccc tgtggagtta ttattaagag
     1021 gcagcagcag aaggctggac ctgctgcacc agcagctgca ggagcttcat gcccacgtgg
     1081 tgctttaagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     1141 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1201 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1321 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1381 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1441 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1501 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1561 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1621 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1681 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     1741 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     1801 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1861 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1921 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     1981 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2041 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2101 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2161 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2221 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2281 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2341 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2401 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2461 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2521 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2581 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2641 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2701 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2761 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2821 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     2881 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2941 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3001 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3061 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3121 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3181 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3241 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3301 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3361 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3421 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3481 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3541 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     3601 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3661 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3721 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3781 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3841 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3901 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3961 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4021 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4081 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4141 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4201 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4261 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4321 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4381 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
