LOCUS       sequence-350953-        6316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddRFP sensor of Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6316)
  AUTHORS   Kim J , Lee S, Jung K, Oh WC, Kim N, Son S, Jo Y, Kwon HB,  Heo WD
  TITLE     Intensiometric biosensors visualize the activity of multiple small 
            GTPases in vivo.
  JOURNAL   Nat Commun. 2019 Jan 14;10(1):211. doi: 10.1038/s41467-018-08217-3.
  PUBMED    30643148
REFERENCE   2  (bases 1 to 6316)
  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: "2019-01-14"; volume: "10(1):211. doi"; pages: " 
            10.1038/s41467-018-08217-3"
COMMENT     Sequence Label: pCMV-R-Rac1
FEATURES             Location/Qualifiers
     source          1..6316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1509..1565
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2349..2927
                     /codon_start=1
                     /product="human Rac family small GTPase 1"
                     /label=Rac1
                     /translation="MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH
                     CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR
                     GLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     misc_feature    2928..2980
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3104..3225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3141..3160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3195..3214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3232..3687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3369..3390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3581..3600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3714..3818
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3820..4177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3820..4011
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3843..3863)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4028..4163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4212..5006
                     /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             4212..5006
                     /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(4266..4285)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4876..4895
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5194..5213)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5238..5285
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5614..6202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6103..6122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      601 tggccaccat caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc
      661 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      721 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
      781 tcatgtacgg ctccgaggcg tacgtgaggc accccgccga catccccgat tacaagaagc
      841 tgcccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaagac ggcggtctgg
      901 tgaccgttac ccaggactcc tccctgcagg acggcacgct gatctgcaag gtgaagatgc
      961 gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1021 cctccaccga gatgctgtac cccgaagacg gcgtgctgaa gggccatagc tatcaggccc
     1081 tgaagctgaa ggacggcggc cactacctgg tggagttcga gaccatctac atggccaaga
     1141 agcccgtgca actgcccggc gattactgtg tggacaccaa gctggacatc acctcccaca
     1201 acgaggacta caccatcgtg gaacagtacg ggcgctccga gggccgccac cgtctgggca
     1261 tggacgagcg gtacaagtta gggcatggta caggtagcac aggcagcggt agctccggta
     1321 ccctgtacaa gggcagcggc agctccattt tacctggaga taaaacaaat aaaaagaaag
     1381 agaaagagcg gccagagatt tctctccctt cagattttga acacacaatt catgtcggtt
     1441 ttgatgctgt cacaggggag tttacgggaa tgccagagca gtgggcccgc ttgcttcagg
     1501 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     1561 gaccaatggt gagcaagagc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgct
     1621 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1681 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     1741 gggacatcct gtccccccag atcatgtacg gctccaaggc gtacgtgaag caccccgccg
     1801 acgtccccga ttacatgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatgc
     1861 acttcgagga cggcggtctg gtgaccgtga ctcaggacac ctccctgcag gacggcacgc
     1921 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     1981 agaagacctt gggctgggat tatgccaccg agcgcctgta ccccgaagac ggcgtgctga
     2041 agggcgagct tctggggcgc ctgaagctga aggacggcgg cctcaacctg gtggagttca
     2101 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactacttc gtggacacca
     2161 agctgggcat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     2221 agggccgcca ccacctgggg catggcaccg gcagcaccgg cagcggcagc tccggtaccg
     2281 cctcctccga ggacaacaac ctgtacaagt ccggactcag atctcgagct caagcttcga
     2341 attctgcaat gcaggccatc aagtgtgtgg tggtgggaga cggagctgta ggtaaaactt
     2401 gcctactgat cagttacaca accaatgcat ttcctggaga atatatccct actgtctttg
     2461 acaattattc tgccaatgtt atggtagatg gaaaaccggt gaatctgggc ttatgggata
     2521 cagctggaca agaagattat gacagattac gccccctatc ctatccgcaa acagatgtgt
     2581 tcttaatttg cttttccctt gtgagtcctg catcatttga aaatgtccgt gcaaagtggt
     2641 atcctgaggt gcggcaccac tgtcccaaca ctcccatcat cctagtggga actaaacttg
     2701 atcttaggga tgataaagac acgatcgaga aactgaagga gaagaagctg actcccatca
     2761 cctatccgca gggtctagcc atggctaagg agattggtgc tgtaaaatac ctggagtgct
     2821 cggcgctcac acagcgaggc ctcaagacag tgtttgacga agcgatccga gcagtcctct
     2881 gcccgcctcc cgtgaagaag aggaagagaa aatgcctgct gttgtaactc gagctcaagc
     2941 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat
     3001 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3061 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3121 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3181 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     3241 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3301 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3361 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3421 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3481 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3541 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3601 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3661 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3721 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3781 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3841 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3901 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3961 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4021 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4081 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4141 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4201 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4261 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4321 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4381 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4441 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4501 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4561 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4621 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4681 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4741 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4801 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4861 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4921 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4981 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5041 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5101 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5161 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5221 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5281 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5341 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5401 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5461 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5521 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5761 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6241 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6301 tattaccgcc atgcat
//
