LOCUS       sequence-346294-        4910 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 4910)
  AUTHORS   Mahlandt EK, Kreider-Letterman G, Chertkova AO, Garcia-Mata R, 
            Goedhart J
  TITLE     Cell-based optimization and characterization of genetically encoded 
            location-based biosensors for Cdc42 or Rac activity.
  JOURNAL   J Cell Sci. 2023 May 15;136(10):jcs260802. doi: 10.1242/jcs.260802. 
            Epub 2023 May 25.
  PUBMED    37226883
REFERENCE   2  (bases 1 to 4910)
  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."; date: "2023-05-15"; volume: "136(10):jcs260802. doi"; pages: 
            " 10.1242/jcs.260802. Epub 2023 May 25"
COMMENT     Sequence Label: HaloTag-2xrGBD
FEATURES             Location/Qualifiers
     source          1..4910
                     /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"
     polyA_signal    1698..1819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1735..1754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1789..1808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1826..2281)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1963..1984)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2175..2194
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2308..2412
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2414..2771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2414..2605
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2437..2457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2622..2757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2806..3600
                     /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             2806..3600
                     /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(2860..2879)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3470..3489
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3788..3807)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3832..3879
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4208..4796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4697..4716
                     /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 ggccgagatc ggcaccggct
      181 tccccttcga cccccactac gtggaggtgc tgggcgagcg catgcactac gttgacgtgg
      241 gcccccgcga cggcaccccc gtgctgttcc tgcacggcaa ccccaccagc agctacgtgt
      301 ggcgcaacat catcccccac gtggccccca cccaccgcac tatcgccccc gacctgatcg
      361 gcatgggcaa gagcgacaag cccgacctgg gctacttctt cgacgaccac gtgcgcttca
      421 tggacgcctt catcgaggcc ctgggcctgg aggaggtggt gctggtgatc cacgactggg
      481 gcagcgccct gggcttccac tgggccaagc gcaaccccga gcgcgtgaag ggcatcgcct
      541 tcatggagtt catccgcccc atccccacct gggacgagtg gcccgagttc gcccgcgaga
      601 ccttccaggc cttccgcacc accgacgtgg gccgcaagct gatcatcgac cagaacgtgt
      661 tcatcgaggg caccctgccc atgggcgtgg tgcgccccct gaccgaggtg gagatggacc
      721 actaccgcga gcccttcctg aaccccgtgg accgcgagcc cctgtggcgc ttccccaacg
      781 agctgcccat cgccggcgag cccgccaaca tcgtggccct ggtggaggag tacatggact
      841 ggctgcacca gagccccgtg cccaagctgc tgttctgggg cacccccggc gtgctgatcc
      901 cccccgccga ggccgcccgc ctggccaaga gcctgcccaa cgttaaggcc gtggacatcg
      961 gccccggcct gaacctgctc caggaggaca accccgacct gatcggcagc gagatcgccc
     1021 gctggctgag caccctggag atcagcggcg tgtacatccc cctcgagtcc atcctggagg
     1081 acctcaatat gctctacatc cggcagatgg cactcagcct ggaggacaca gagctgcaga
     1141 ggaaactaga tcatgagatc cggatgaggg atggggcctg caagctgctg gcagcctgct
     1201 cccagcgaga gcaggctctg gaagccacca agagcctgct ggtgtgcaac agccgtattc
     1261 tcagctacat gggtgagctg cagcggcgaa aggaggccca ggtgctggag aagacaggca
     1321 ggcgtcgagt tatcctggag gacctcaata tgctctacat ccggcagatg gcactcagcc
     1381 tggaggacac agagctgcag aggaaactag atcatgagat ccggatgagg gatggggcct
     1441 gcaagctgct ggcagcctgc tcccagcgag agcaggctct ggaagccacc aagagcctgc
     1501 tggtgtgcaa cagccgtatt ctcagctaca tgggtgagct gcagcggcga aaggaggccc
     1561 aggtgctgga gaagacaggc tctagataac tgatcataat cagccatacc acatttgtag
     1621 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1681 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1741 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1801 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1861 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1921 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1981 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2041 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2101 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2161 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2221 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2401 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2521 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2581 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2641 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2701 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2761 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2821 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2881 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2941 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3001 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3061 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3121 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3181 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3241 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3301 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3361 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3421 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3481 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3541 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3601 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3661 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3721 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3781 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3841 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3901 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3961 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4021 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4321 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
