LOCUS       sequence-355542-        6949 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 6949)
  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 6949)
  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: pAAV-CAG-G-Rac1
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..403
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     465..484
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1437..1493
                     /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             2259..2837
                     /codon_start=1
                     /product="human Rac family small GTPase 1"
                     /label=Rac1
                     /translation="MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH
                     CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR
                     GLKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     misc_feature    2846..3434
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2899..2919)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3317..3328)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3477..3598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3488..3507)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3542..3561
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3760..3889
                     /label=AAV2 ITR
     rep_origin      complement(4126..4714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4206..4225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4885..5676,5677..5745))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5508..5527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5746..5850)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5876..6331)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6013..6034)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6225..6244
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6403..6532
                     /label=AAV2 ITR
ORIGIN
        1 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
       61 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      121 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      181 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      241 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      301 gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg
      361 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
      421 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
      481 tgtgctgtct catcattttg gcaaagaatt gctcgagcgc tagcaccatg gccaccatca
      541 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
      601 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
      661 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
      721 ccgaggcgta cgtgaggcac cccgccgaca tccccgatta caagaagctg cccttccccg
      781 agggcttcaa gtgggagcgc gtgatgaact tcgaagacgg cggtctggtg accgttaccc
      841 aggactcctc cctgcaggac ggcacgctga tctgcaaggt gaagatgcgc ggcaccaact
      901 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgaga
      961 tgctgtaccc cgaagacggc gtgctgaagg gccatagcta tcaggccctg aagctgaagg
     1021 acggcggcca ctacctggtg gagttcgaga ccatctacat ggccaagaag cccgtgcaac
     1081 tgcccggcga ttactgtgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     1141 ccatcgtgga acagtacggg cgctccgagg gccgccaccg tctgggcatg gacgagcggt
     1201 acaagttagg gcatggtaca ggtagcacag gcagcggtag ctccggtacc ctgtacaagg
     1261 gcggcggcag ctccatttta cctggagata aaacaaataa aaagaaagag aaagagcggc
     1321 cagagatttc tctcccttca gattttgaac acacaattca tgtcggtttt gatgctgtca
     1381 caggggagtt tacgggaatg ccagagcagt gggcccgctt gcttcaggga tccggcgcaa
     1441 caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccaatggtga
     1501 gcaagagcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcttg gagggctcca
     1561 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccttac gagggcaccc
     1621 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
     1681 cccccctgat catgtacggc tccaagatgt acgtgaagca ccccgccgac gtccccgatt
     1741 acatgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgcac ttcgaggacg
     1801 gcggtctggt gaccgcgaca caggactcct ccctgcagga cggcacgctg atctacaagg
     1861 tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccttgg
     1921 gctgggatta tgccaccgag cgcctgtacc ccgaagaagg cgtgctgaag ggcgagctcc
     1981 tggggcgcct gaagctgaag gacggcggcc tcaacctggt ggagtccaag accatctaca
     2041 tggccaagaa gcccgtgcaa ctgcccggct actacttcgt ggacaccaag ctggacatca
     2101 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc
     2161 acctggggca tggtacaggt agcacaggca gcggtagctc cggtaccctg tacaagtccg
     2221 gactcagatc tcgagctcaa gcttcgaatt ctgccaccat gcaggccatc aagtgtgtgg
     2281 tggtgggaga cggagctgta ggtaaaactt gcctactgat cagttacaca accaatgcat
     2341 ttcctggaga atatatccct actgtctttg acaattattc tgccaatgtt atggtagatg
     2401 gaaaaccggt gaatctgggc ttatgggata cagctggaca agaagattat gacagattac
     2461 gccccctatc ctatccgcaa acagatgtgt tcttaatttg cttttccctt gtgagtcctg
     2521 catcatttga aaatgtccgt gcaaagtggt atcctgaggt gcggcaccac tgtcccaaca
     2581 ctcccatcat cctagtggga actaaacttg atcttaggga tgataaagac acgatcgaga
     2641 aactgaagga gaagaagctg actcccatca cctatccgca gggtctagcc atggctaagg
     2701 agattggtgc tgtaaaatac ctggagtgct cggcgctcac acagcgaggc ctcaagacag
     2761 tgtttgacga agcgatccga gcagtcctct gcccgcctcc cgtgaagaag aggaagagaa
     2821 aatgcctgct gttgtaaatc tccggaatca acctctggat tacaaaattt gtgaaagatt
     2881 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2941 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3001 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3061 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3121 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3181 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3241 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3301 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3361 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3421 ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag
     3481 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3541 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3601 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3661 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     3721 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     3781 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3841 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg
     3901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5881 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5941 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6001 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6061 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6121 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6181 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6241 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6301 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     6361 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc
     6421 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     6481 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta
     6541 atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagag taccattgac
     6601 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6661 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6721 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     6781 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6841 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     6901 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcg
//
