LOCUS       sequence-355462-        6323 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 6323)
  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 6323)
  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..6323
                     /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"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1521..1577
                     /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             2343..2921
                     /codon_start=1
                     /product="human Rac family small GTPase 1"
                     /label=Rac1
                     /translation="MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH
                     CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR
                     GLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     misc_feature    2922..2987
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3111..3232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3202..3221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3239..3694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3376..3397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3588..3607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3721..3825
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3827..4184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3827..4018
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3850..3870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4035..4170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4219..5013
                     /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             4219..5013
                     /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(4273..4292)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4883..4902
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5201..5220)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5245..5292
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5621..6209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6110..6129
                     /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 ccggtgccac catggccacc atcaaagagt tcatgcgctt caaggtgcgc atggagggct
      661 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      721 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
      781 tgtcccccca gttcatgtac ggctccgagg cgtacgtgag gcaccccgcc gacatccccg
      841 attacaagaa gctgcccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgaag
      901 acggcggtct ggtgaccgtt acccaggact cctccctgca ggacggcacg ctgatctgca
      961 aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca
     1021 tgggctggga ggcctccacc gagatgctgt accccgaaga cggcgtgctg aagggccata
     1081 gctatcaggc cctgaagctg aaggacggcg gccactacct ggtggagttc gagaccatct
     1141 acatggccaa gaagcccgtg caactgcccg gcgattactg tgtggacacc aagctggaca
     1201 tcacctccca caacgaggac tacaccatcg tggaacagta cgggcgctcc gagggccgcc
     1261 accgtctggg catggacgag cggtacaagt tagggcatgg tacaggtagc acaggcagcg
     1321 gtagctccgg taccctgtac aagggcagcg gcagctccat tttacctgga gataaaacaa
     1381 ataaaaagaa agagaaagag cggccagaga tttctctccc ttcagatttt gaacacacaa
     1441 ttcatgtcgg ttttgatgct gtcacagggg agtttacggg aatgccagag cagtgggccc
     1501 gcttgcttca gggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg
     1561 aagagaatcc tggaccgatg gtgagcaaga gcgaggaggt catcaaagag ttcatgcgct
     1621 tcaaggtgcg cttggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1681 agggccgccc ttacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     1741 tgcccttcgc ctgggacatc ctgtcccccc tgatcatgta cggctccaag atgtacgtga
     1801 agcaccccgc cgacgtcccc gattacatga agctgtcctt ccccgagggc ttcaagtggg
     1861 agcgcgtgat gcacttcgag gacggcggtc tggtgaccgc gacacaggac tcctccctgc
     1921 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     1981 ccgtaatgca gaagaagacc ttgggctggg attatgccac cgagcgcctg taccccgaag
     2041 aaggcgtgct gaagggcgag ctcctggggc gcctgaagct gaaggacggc ggcctcaacc
     2101 tggtggagtc caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     2161 tcgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2221 acgagcgctc cgagggccgc caccacctgg ggcatggtac aggtagcaca ggcagcggta
     2281 gctccggtac cctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcca
     2341 ccatgcaggc catcaagtgt gtggtggtgg gagacggagc tgtaggtaaa acttgcctac
     2401 tgatcagtta cacaaccaat gcatttcctg gagaatatat ccctactgtc tttgacaatt
     2461 attctgccaa tgttatggta gatggaaaac cggtgaatct gggcttatgg gatacagctg
     2521 gacaagaaga ttatgacaga ttacgccccc tatcctatcc gcaaacagat gtgttcttaa
     2581 tttgcttttc ccttgtgagt cctgcatcat ttgaaaatgt ccgtgcaaag tggtatcctg
     2641 aggtgcggca ccactgtccc aacactccca tcatcctagt gggaactaaa cttgatctta
     2701 gggatgataa agacacgatc gagaaactga aggagaagaa gctgactccc atcacctatc
     2761 cgcagggtct agccatggct aaggagattg gtgctgtaaa atacctggag tgctcggcgc
     2821 tcacacagcg aggcctcaag acagtgtttg acgaagcgat ccgagcagtc ctctgcccgc
     2881 ctcccgtgaa gaagaggaag agaaaatgcc tgctgttgta atccggactc agatctcgag
     2941 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat
     3001 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3061 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3121 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3181 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     3241 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3301 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3361 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3421 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3481 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3541 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3601 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3661 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3841 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4021 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4081 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4141 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4201 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4261 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4321 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4381 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4441 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4501 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4561 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4621 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4681 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4741 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4801 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4861 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4921 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4981 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5041 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5101 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5161 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5221 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5281 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5341 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5401 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5461 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5521 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5761 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6241 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6301 ataaccgtat taccgccatg cat
//
