LOCUS       sequence-429691-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rac activity sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6120)
  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 6120)
  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-3x-p67phox-GBD
FEATURES             Location/Qualifiers
     source          1..6120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(91..546)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(228..249)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     440..459
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        573..677
                     /gene="bla"
                     /label=AmpR promoter
     promoter        679..1036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        679..870
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(702..722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      887..1022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1071..1865
                     /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             1071..1865
                     /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(1125..1144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1735..1754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2053..2072)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2097..2144
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2473..3061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2962..2981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      3339..3348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3345..4052
                     /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(3490..3508)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3758..3777
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    join(6083..6120,1..84)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(6120,1..19))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
       61 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
      121 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
      181 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
      241 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
      301 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
      361 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
      421 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
      481 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
      541 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      601 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      661 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
      721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
      841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
      901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
      961 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     1021 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     1081 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     1141 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     1201 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     1261 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     1321 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     1381 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     1441 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     1501 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     1561 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     1621 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     1681 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     1741 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     1801 acggtatcgc cgctcccgat tcacagcgca tcgccttcta tcgccttctt gacgagttct
     1861 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     1921 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     1981 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     2041 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     2101 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     2161 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     2221 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     2281 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     2341 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2401 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2581 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2641 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2701 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2761 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2821 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2881 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2941 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3001 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3061 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctggcct
     3121 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3181 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     3241 gttccgcgtt acataactta cggtaaatgg cccgcctggg aggtctatat aagcagagct
     3301 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
     3361 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     3421 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3481 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     3541 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     3601 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3661 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     3721 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     3781 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     3841 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     3901 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     3961 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     4021 actccaccgg cggcatggac gagctgtaca agtccggact cagatctcga gctcacatgt
     4081 ccctggtgga ggccatcagc ctctggaatg aaggggtgct ggcagcggac aagaaggact
     4141 ggaagggagc cctggatgcc ttcagtgccg tccaggaccc ccactcccgg atttgcttca
     4201 acattggctg catgtacact atcctgaaga acatgactga agcagagaag gcctttacca
     4261 gaagcattaa ccgagacaag cacttggcag tggcttactt ccaacgaggg atgctctact
     4321 accagacaga gaaatatgat ttggctatca aagaccttaa agaagccttg attcagcttc
     4381 gagggaatca gctgatagac tataagatcc tggggctcca gttcaagctg tttgcctgtg
     4441 aggtgttata taacattgct ttcatgtatg ccaagaagga ggaatggaaa aaagctgaag
     4501 aacagttagc attggccacg agcatgaagt ctgagcccag acattccaaa atcgacaagg
     4561 cgatggagtg tgtctggaag cagaagctat atgagccagt ggtgatccct gtgggcaagc
     4621 tgtttcgacc aaatgagaga caagtggctc agctggccaa gaaggattac ctaggcaagg
     4681 cgacgggttc tggaagtgga tcggttctca tgtccctggt ggaggccatc agcctctgga
     4741 atgaaggggt gctggcagcg gacaagaagg actggaaggg agccctggat gccttcagtg
     4801 ccgtccagga cccccactcc cggatttgct tcaacattgg ctgcatgtac actatcctga
     4861 agaacatgac tgaagcagag aaggccttta ccagaagcat taaccgagac aagcacttgg
     4921 cagtggctta cttccaacga gggatgctct actaccagac agagaaatat gatttggcta
     4981 tcaaagacct taaagaagcc ttgattcagc ttcgagggaa tcagctgata gactataaga
     5041 tcctggggct ccagttcaag ctgtttgcct gtgaggtgtt atataacatt gctttcatgt
     5101 atgccaagaa ggaggaatgg aaaaaagctg aagaacagtt agcattggcc acgagcatga
     5161 agtctgagcc cagacattcc aaaatcgaca aggcgatgga gtgtgtctgg aagcagaagc
     5221 tatatgagcc agtggtgatc cctgtgggca agctgtttcg accaaatgag agacaagtgg
     5281 ctcagctggc caagaaggat tacctaggca aggcgacggg ttctggaagt ggatcgaatt
     5341 ccatgtccct ggtggaggcc atcagcctct ggaatgaagg ggtgctggca gcggacaaga
     5401 aggactggaa gggagccctg gatgccttca gtgccgtcca ggacccccac tcccggattt
     5461 gcttcaacat tggctgcatg tacactatcc tgaagaacat gactgaagca gagaaggcct
     5521 ttaccagaag cattaaccga gacaagcact tggcagtggc ttacttccaa cgagggatgc
     5581 tctactacca gacagagaaa tatgatttgg ctatcaaaga ccttaaagaa gccttgattc
     5641 agcttcgagg gaatcagctg atagactata agatcctggg gctccagttc aagctgtttg
     5701 cctgtgaggt gttatataac attgctttca tgtatgccaa gaaggaggaa tggaaaaaag
     5761 ctgaagaaca gttagcattg gccacgagca tgaagtctga gcccagacat tccaaaatcg
     5821 acaaggcgat ggagtgtgtc tggaagcaga agctatatga gccagtggtg atccctgtgg
     5881 gcaagctgtt tcgaccaaat gagagacaag tggctcagct ggccaagaag gattacctag
     5941 gcaaggcgac gtagggatcc atggatctag ataactgatc ataatcagcc ataccacatt
     6001 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6061 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
//
