LOCUS       sequence-346255-        4979 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 4979)
  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 4979)
  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-3xrGBD
FEATURES             Location/Qualifiers
     source          1..4979
                     /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    1767..1888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1804..1823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1858..1877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1895..2350)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2032..2053)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2244..2263
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2377..2481
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2483..2840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2483..2674
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2506..2526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2691..2826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2875..3669
                     /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             2875..3669
                     /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(2929..2948)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3539..3558
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3857..3876)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3901..3948
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4277..4865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4766..4785
                     /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 gtcgagtcca
     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 agacaggcag gcgtcgagtt atcctggagg acctcaatat gctctacatc cggcagatgg
     1441 cactcagcct ggaggacaca gagctgcaga ggaaactaga tcatgagatc cggatgaggg
     1501 atggggcctg caagctgctg gcagcctgct cccagcgaga gcaggctctg gaagccacca
     1561 agagcctgct ggtgtgcaac agccgtattc tcagctacat gggtgagctg cagcggcgaa
     1621 aggaggccca ggtgctggag aagacaggct ctagataact gatcataatc agccatacca
     1681 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1741 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1861 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     1921 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1981 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2041 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2101 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2161 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2221 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2281 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2341 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2401 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2461 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2521 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2581 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2641 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2701 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2761 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2821 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2881 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2941 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3001 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3061 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3121 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3181 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3241 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3301 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3361 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3421 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3481 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3541 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3601 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3661 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3721 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3781 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3841 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3901 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     3961 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4021 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4081 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
