LOCUS       sequence-429695-        4776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rho activity sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4776)
  AUTHORS   Nanda S, Calderon A, Sachan A, Duong TT, Koch J, Xin X, 
            Solouk-Stahlberg D, Wu YW, Nalbant P, Dehmelt L
  TITLE     Rho GTPase activity crosstalk mediated by Arhgef11 and Arhgef12 
            coordinates cell protrusion-retraction cycles.
  JOURNAL   Nat Commun. 2023 Dec 15;14(1):8356. doi: 10.1038/s41467-023-43875-y.
  PUBMED    38102112
REFERENCE   2  (bases 1 to 4776)
  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: "Nat 
            Commun."; date: "2023-12-15"; volume: "14(1):8356. doi"; pages: " 
            10.1038/s41467-023-43875-y"
COMMENT     Sequence Label: delCMV-mCherry-2x-Rhotekin-GBD
FEATURES             Location/Qualifiers
     source          1..4776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    97..218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(134..153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     188..207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(225..680)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(362..383)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     574..593
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        707..811
                     /gene="bla"
                     /label=AmpR promoter
     promoter        813..1170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        813..1004
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(836..856)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1021..1156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1205..1999
                     /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             1205..1999
                     /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(1259..1278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1869..1888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2187..2206)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2231..2278
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2607..3195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3096..3115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      3473..3482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3479..4186
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3624..3642)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3892..3911
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
       61 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
      121 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
      181 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
      241 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      301 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      361 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
      421 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
      481 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
      541 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
      601 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
      661 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
      721 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      781 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
      841 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
      901 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      961 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     1021 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     1081 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     1141 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     1201 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     1261 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     1321 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     1381 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     1441 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     1501 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     1561 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     1621 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1681 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     1741 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1801 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1861 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1921 cttcctcgtg ctttacggta tcgccgctcc cgattcacag cgcatcgcct tctatcgcct
     1981 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     2041 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     2101 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     2161 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     2221 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     2281 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     2341 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     2401 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     2461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3301 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     3361 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tgggaggtct
     3421 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
     3481 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3541 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3601 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3661 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3721 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3781 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3841 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3901 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3961 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     4021 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     4081 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     4141 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc
     4201 tcgagctcaa gcttcgaatt ccctggagga cctcaatatg ctctacatcc ggcagatggc
     4261 actcagcctg gaggacacag agctgcagag gaaactagat catgagatcc ggatgaggga
     4321 tggggcctgc aagctgctgg cagcctgctc ccagcgagag caggctctgg aagccaccaa
     4381 gagcctgctg gtgtgcaaca gccgtattct cagctacatg ggtgagctgc agcggcgaaa
     4441 ggaggcccag gtgctggaga agacaggctc cggactcgct caagcttcga attccctgga
     4501 ggacctcaat atgctctaca tccggcagat ggcactcagc ctggaggaca cagagctgca
     4561 gaggaaacta gatcatgaga tccggatgag ggatggggcc tgcaagctgc tggcagcctg
     4621 ctcccagcga gagcaggctc tggaagccac caagagcctg ctggtgtgca acagccgtat
     4681 tctcagctac atgggtgagc tgcagcggcg aaaggaggcc caggtgctgg agaagacagg
     4741 ctagggatcc atggatctag ataactgatc ataatc
//
