LOCUS       sequence-355545-        6320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddRFP sensor of Cdc42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6320)
  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 6320)
  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-Cdc42
FEATURES             Location/Qualifiers
     source          1..6320
                     /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             1516..1572
                     /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"
     misc_feature    2932..2984
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3108..3229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3145..3164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3199..3218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3236..3691)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3373..3394)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3585..3604
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3718..3822
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3824..4181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3824..4015
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3847..3867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4032..4167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4216..5010
                     /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             4216..5010
                     /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(4270..4289)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4880..4899
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5198..5217)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5242..5289
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5618..6206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6107..6126
                     /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 gctagcgcta
      601 agcaccatgg ccaccatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg
      661 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      721 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
      781 ccccagttca tgtacggctc cgaggcgtac gtgaggcacc ccgccgacat ccccgattac
      841 aagaagctgc ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaagacggc
      901 ggtctggtga ccgttaccca ggactcctcc ctgcaggacg gcacgctgat ctgcaaggtg
      961 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     1021 tgggaggcct ccaccgagat gctgtacccc gaagacggcg tgctgaaggg ccatagctat
     1081 caggccctga agctgaagga cggcggccac tacctggtgg agttcgagac catctacatg
     1141 gccaagaagc ccgtgcaact gcccggcgat tactgtgtgg acaccaagct ggacatcacc
     1201 tcccacaacg aggactacac catcgtggaa cagtacgggc gctccgaggg ccgccaccgt
     1261 ctgggcatgg acgagcggta caagttaggg catggtacag gtagcacagg cagcggtagc
     1321 tccggtaccc tgtacaaggg cagcggcagc tccattttac ctggagataa aacaaataaa
     1381 aagaaagaga aagagcggcc agagatttct ctcccttcag attttgaaca cacaattcat
     1441 gtcggttttg atgctgtcac aggggagttt acgggaatgc cagagcagtg ggcccgcttg
     1501 cttcagggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     1561 aaccctggac caatggtgag caagagcgag gaggtcatca aagagttcat gcgcttcaag
     1621 gtgcgcttgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1681 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     1741 ttcgcctggg acatcctgtc cccccagatc atgtacggct ccaaggcgta cgtgaagcac
     1801 cccgccgacg tccccgatta catgaagctg tccttccccg agggcttcaa gtgggagcgc
     1861 gtgatgcact tcgaggacgg cggtctggtg accgtgactc aggacacctc cctgcaggac
     1921 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     1981 atgcagaaga agaccttggg ctgggattat gccaccgagc gcctgtaccc cgaagacggc
     2041 gtgctgaagg gcgagcttct ggggcgcctg aagctgaagg acggcggcct caacctggtg
     2101 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctacttcgtg
     2161 gacaccaagc tgggcatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     2221 cgctccgagg gccgccacca cctggggcat ggcaccggca gcaccggcag cggcagctcc
     2281 ggtaccgcct cctccgagga caacaacctg tacaagtccg gactcagatc tcgagctcaa
     2341 gcttcgaatt cggcaatgca gacaattaag tgtgttgttg tgggcgatgg tgctgttggt
     2401 aaaacatgtc tcctgatatc ctacacaaca aacaaatttc catcggaata tgtaccgact
     2461 gtttttgaca actatgcagt cacagttatg attggtggag aaccatatac tcttggactt
     2521 tttgatactg cagggcaaga ggattatgac agattacgac cgctgagtta tccacaaaca
     2581 gatgtatttc tagtctgttt ttcagtggtc tctccatctt catttgaaaa cgtgaaagaa
     2641 aagtgggtgc ctgagataac tcaccactgt ccaaagactc ctttcttgct tgttgggact
     2701 caaattgatc tcagagatga cccctctact attgagaaac ttgccaagaa caaacagaag
     2761 cctatcactc cagagactgc tgaaaagctg gcccgtgacc tgaaggctgt caagtatgtg
     2821 gagtgttctg cacttacaca gagaggtctg aagaatgtgt ttgatgaggc tatcctagct
     2881 gccctcgagc ctccggaaac tcaacccaaa aggaagtgct gtatattcta actcgagctc
     2941 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
     3001 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3061 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3121 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3181 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3241 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3301 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3361 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3421 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3481 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3541 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3601 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3661 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3721 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3781 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3841 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3901 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3961 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4021 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4081 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4141 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4201 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4261 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4321 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4381 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4441 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4501 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4561 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4621 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4681 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4741 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4801 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4861 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4921 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4981 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5041 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5101 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5161 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5221 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5281 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5341 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5401 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5461 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5521 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5581 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5641 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5701 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5761 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta tcaccgccta
     5821 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5881 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5941 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6001 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6061 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6301 accgtattac cgccatgcat
//
