LOCUS       sequence-355653-        4096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP sensor of Hras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4096)
  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 4096)
  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-Hras
FEATURES             Location/Qualifiers
     source          1..4096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     380..402
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     464..483
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1534..1590
                     /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             2314..2883
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=H-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     misc_feature    2892..3480
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2945..2965)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3363..3374)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3523..3644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3534..3553)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3588..3607
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3806..3935
                     /label=AAV2 ITR
ORIGIN
        1 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
       61 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      121 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      181 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      241 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
      301 agggcccttt gtgcgggggg agcggctcgg ggctgtccgc ggggggacgg ctgccttcgg
      361 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
      421 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
      481 gtgctgtctc atcattttgg caaagaattg ctcgaggtca ccatggccac catcaaagag
      541 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag
      601 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
      661 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccgag
      721 gcgtacgtga ggcaccccgc cgacatcccc gattacaaga agctgccctt ccccgagggc
      781 ttcaagtggg agcgcgtgat gaacttcgaa gacggcggtc tggtgaccgt tacccaggac
      841 tcctccctgc aggacggcac gctgatctgc aaggtgaaga tgcgcggcac caacttcccc
      901 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagatgctg
      961 taccccgaag acggcgtgct gaagggccat agctatcagg ccctgaagct gaaggacggc
     1021 ggccactacc tggtggagtt cgagaccatc tacatggcca agaagcccgt gcaactgccc
     1081 ggcgattact gtgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     1141 gtggaacagt acgggcgctc cgagggccgc caccgtctgg gcatggacga gcggtacaag
     1201 ttagggcatg gtacaggtag cacaggcagc ggtagctccg gtaccctgta caagtccgga
     1261 ctcagatctc gagccaccat gccttctaag acaagcaaca ctatccgtgt tttcttgccg
     1321 aacaagcaaa gaacagtggt caatgtgcga aatggaatga gcttgcatga ctgccttatg
     1381 aaagcactca aggtgagggg cctgcaacca gagtgctgtg cagtgttcag acttctccac
     1441 gaacacaaag gtaaaaaagc acgcttagat tggaatactg atgctgcgtc tttgattgga
     1501 gaagaacttc aagtagattt cctgggatcc ggcgcaacaa acttctctct gctgaaacaa
     1561 gccggagatg tcgaagagaa tcctggaccg atggtgagca agagcgagga ggtcatcaaa
     1621 gagttcatgc gcttcaaggt gcgcttggag ggctccatga acggccacga gttcgagatc
     1681 gagggcgagg gcgagggccg cccttacgag ggcacccaga ccgccaagct gaaggtgacc
     1741 aagggcggcc ccctgccctt cgcctgggac atcctgtccc ccctgatcat gtacggctcc
     1801 aagatgtacg tgaagcaccc cgccgacgtc cccgattaca tgaagctgtc cttccccgag
     1861 ggcttcaagt gggagcgcgt gatgcacttc gaggacggcg gtctggtgac cgcgacacag
     1921 gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc
     1981 ccccccgacg gccccgtaat gcagaagaag accttgggct gggattatgc caccgagcgc
     2041 ctgtaccccg aagaaggcgt gctgaagggc gagctcctgg ggcgcctgaa gctgaaggac
     2101 ggcggcctca acctggtgga gtccaagacc atctacatgg ccaagaagcc cgtgcaactg
     2161 cccggctact acttcgtgga caccaagctg gacatcacct cccacaacga ggactacacc
     2221 atcgtggaac agtacgagcg ctccgagggc cgccaccacc tggggcatgg tacaggtagc
     2281 acaggcagcg gtagctccgg taccctgtac aagatgacgg aatataagct ggtggtggtg
     2341 ggcgccggcg gtgtgggcaa gagtgcgctg accatccagc tgatccagaa ccattttgtg
     2401 gacgaatacg accccactat agaggattcc taccggaagc aggtggtcat tgatggggag
     2461 acgtgcctgt tggacatcct ggataccgcc ggccaggagg agtacagcgc catgcgggac
     2521 cagtacatgc gcaccgggga gggcttcctg tgtgtgtttg ccatcaacaa caccaagtct
     2581 tttgaggaca tccaccagta cagggagcag atcaaacggg tgaaggactc ggatgacgtg
     2641 cccatggtgc tggtggggaa caagtgtgac ctggctgcac gcactgtgga atctcggcag
     2701 gctcaggacc tcgcccgaag ctacggcatc ccctacatcg agacctcggc caagacccgg
     2761 cagggagtgg aggatgcctt ctacacgttg gtgcgtgaga tccggcagca caagctgcgg
     2821 aagctgaacc ctcctgatga gagtggcccc ggctgcatga gctgcaagtg tgtgctctcc
     2881 tgaatctccg gaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2941 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3001 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3061 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3121 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3181 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3241 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3301 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3361 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3421 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3481 atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     3541 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3601 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3661 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3721 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     3781 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     3841 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3901 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg
     3961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4081 ccacagaatc agggga
//
