LOCUS       sequence-350952-        6322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP sensor of Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6322)
  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 6322)
  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-G-Rac1
FEATURES             Location/Qualifiers
     source          1..6322
                     /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             1530..1586
                     /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             join(2355..2402,2403..2405,2406..2933)
                     /codon_start=1
                     /product="dominant negative mutant of human Rac family 
                     small GTPase 1"
                     /label=Rac1 (T17N)
                     /translation="MQAIKCVVVGDGAVGKNCLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH
                     CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR
                     GLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     CDS             2355..2933
                     /codon_start=1
                     /product="human Rac family small GTPase 1"
                     /label=Rac1
                     /translation="MQAIKCVVVGDGAVGKNCLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH
                     CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR
                     GLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     misc_feature    2934..2986
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3110..3231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3147..3166)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3201..3220
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3238..3693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3375..3396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3587..3606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3720..3824
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3826..4183
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3826..4017
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3849..3869)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4034..4169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4218..5012
                     /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             4218..5012
                     /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(4272..4291)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4882..4901
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5200..5219)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5244..5291
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5620..6208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6109..6128
                     /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 gtcgtacaag tccggactca gatctcgagg cagctccatt ttacctggag
     1381 ataaaacaaa taaaaagaaa gagaaagagc ggccagagat ttctctccct tcagattttg
     1441 aacacacaat tcatgtcggt tttgatgctg tcacagggga gtttacggga atgccagagc
     1501 agtgggcccg cttgcttcag ggaagcggag ctactaactt cagcctgctg aagcaggctg
     1561 gagacgtgga ggagaaccct ggaccaatgg tgagcaagag cgaggaggtc atcaaagagt
     1621 tcatgcgctt caaggtgcgc ttggagggct ccatgaacgg ccacgagttc gagatcgagg
     1681 gcgagggcga gggccgccct tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1741 gcggccccct gcccttcgcc tgggacatcc tgtcccccct gatcatgtac ggctccaaga
     1801 tgtacgtgaa gcaccccgcc gacgtccccg attacatgaa gctgtccttc cccgagggct
     1861 tcaagtggga gcgcgtgatg cacttcgagg acggcggtct ggtgaccgcg acacaggact
     1921 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     1981 ccgacggccc cgtaatgcag aagaagacct tgggctggga ttatgccacc gagcgcctgt
     2041 accccgaaga aggcgtgctg aagggcgagc tcctggggcg cctgaagctg aaggacggcg
     2101 gcctcaacct ggtggagtcc aagaccatct acatggccaa gaagcccgtg caactgcccg
     2161 gctactactt cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     2221 tggaacagta cgagcgctcc gagggccgcc accacctggg gcatggtaca ggtagcacag
     2281 gcagcggtag ctccggtacc ctgtacaagg gcagtggttc cggactcaga tctcgagctc
     2341 aagcttcgaa ttccatgcag gccatcaagt gtgtggtggt gggagacgga gctgtaggta
     2401 aaaattgcct actgatcagt tacacaacca atgcatttcc tggagaatat atccctactg
     2461 tctttgacaa ttattctgcc aatgttatgg tagatggaaa accggtgaat ctgggcttat
     2521 gggatacagc tggacaagaa gattatgaca gattacgccc cctatcctat ccgcaaacag
     2581 atgtgttctt aatttgcttt tcccttgtga gtcctgcatc atttgaaaat gtccgtgcaa
     2641 agtggtatcc tgaggtgcgg caccactgtc ccaacactcc catcatccta gtgggaacta
     2701 aacttgatct tagggatgat aaagacacga tcgagaaact gaaggagaag aagctgactc
     2761 ccatcaccta tccgcagggt ctagccatgg ctaaggagat tggtgctgta aaatacctgg
     2821 agtgctcggc gctcacacag cgaggcctca agacagtgtt tgacgaagcg atccgagcag
     2881 tcctctgccc gcctcccgtg aagaagagga agagaaaatg cctgctgttg taactcgagc
     2941 tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata
     3001 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3061 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3121 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3181 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     3241 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3361 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3421 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3481 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3541 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3601 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3661 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3721 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3781 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3841 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3901 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3961 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4021 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4081 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4141 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4201 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4261 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4321 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4381 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4441 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4501 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4561 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4621 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4681 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4741 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4801 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4861 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4921 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4981 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5041 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5101 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5161 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5221 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5281 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5341 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5401 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5461 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5521 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5581 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5641 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5701 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5761 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5821 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5881 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5941 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6001 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6061 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6121 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6181 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6241 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6301 taaccgtatt accgccatgc at
//
