LOCUS       sequence-355446-        6370 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 6370)
  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 6370)
  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-Hras
FEATURES             Location/Qualifiers
     source          1..6370
                     /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             1620..1676
                     /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             2400..2969
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=H-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     misc_feature    2978..3034
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3158..3279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3195..3214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3249..3268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3286..3741)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3423..3444)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3635..3654
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3768..3872
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3874..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3874..4065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4266..5060
                     /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             4266..5060
                     /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(4320..4339)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4930..4949
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5248..5267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5292..5339
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5668..6256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6157..6176
                     /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 ccggtaccat ggccaccatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca
      661 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
      721 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
      781 ccccccagtt catgtacggc tccgaggcgt acgtgaggca ccccgccgac atccccgatt
      841 acaagaagct gcccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaagacg
      901 gcggtctggt gaccgttacc caggactcct ccctgcagga cggcacgctg atctgcaagg
      961 tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg
     1021 gctgggaggc ctccaccgag atgctgtacc ccgaagacgg cgtgctgaag ggccatagct
     1081 atcaggccct gaagctgaag gacggcggcc actacctggt ggagttcgag accatctaca
     1141 tggccaagaa gcccgtgcaa ctgcccggcg attactgtgt ggacaccaag ctggacatca
     1201 cctcccacaa cgaggactac accatcgtgg aacagtacgg gcgctccgag ggccgccacc
     1261 gtctgggcat ggacgagcgg tacaagttag ggcatggtac aggtagcaca ggcagcggta
     1321 gctccggtac cctgtacaag tccggactca gatctcgagc caccatgcct tctaagacaa
     1381 gcaacactat ccgtgttttc ttgccgaaca agcaaagaac agtggtcaat gtgcgaaatg
     1441 gaatgagctt gcatgactgc cttatgaaag cactcaaggt gaggggcctg caaccagagt
     1501 gctgtgcagt gttcagactt ctccacgaac acaaaggtaa aaaagcacgc ttagattgga
     1561 atactgatgc tgcgtctttg attggagaag aacttcaagt agatttcctg ggatccggcg
     1621 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgg
     1681 tgagcaagag cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc ttggagggct
     1741 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgccct tacgagggca
     1801 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     1861 tgtcccccct gatcatgtac ggctccaaga tgtacgtgaa gcaccccgcc gacgtccccg
     1921 attacatgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg cacttcgagg
     1981 acggcggtct ggtgaccgcg acacaggact cctccctgca ggacggcacg ctgatctaca
     2041 aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacct
     2101 tgggctggga ttatgccacc gagcgcctgt accccgaaga aggcgtgctg aagggcgagc
     2161 tcctggggcg cctgaagctg aaggacggcg gcctcaacct ggtggagtcc aagaccatct
     2221 acatggccaa gaagcccgtg caactgcccg gctactactt cgtggacacc aagctggaca
     2281 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc
     2341 accacctggg gcatggtaca ggtagcacag gcagcggtag ctccggtacc ctgtacaaga
     2401 tgacggaata taagctggtg gtggtgggcg ccggcggtgt gggcaagagt gcgctgacca
     2461 tccagctgat ccagaaccat tttgtggacg aatacgaccc cactatagag gattcctacc
     2521 ggaagcaggt ggtcattgat ggggagacgt gcctgttgga catcctggat accgccggcc
     2581 aggaggagta cagcgccatg cgggaccagt acatgcgcac cggggagggc ttcctgtgtg
     2641 tgtttgccat caacaacacc aagtcttttg aggacatcca ccagtacagg gagcagatca
     2701 aacgggtgaa ggactcggat gacgtgccca tggtgctggt ggggaacaag tgtgacctgg
     2761 ctgcacgcac tgtggaatct cggcaggctc aggacctcgc ccgaagctac ggcatcccct
     2821 acatcgagac ctcggccaag acccggcagg gagtggagga tgccttctac acgttggtgc
     2881 gtgagatccg gcagcacaag ctgcggaagc tgaaccctcc tgatgagagt ggccccggct
     2941 gcatgagctg caagtgtgtg ctctcctgac cggactcaga tctcgagctc aagcttcgaa
     3001 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     3061 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3121 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     3301 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3361 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3421 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3481 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3541 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3601 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3661 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3721 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3781 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3841 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4261 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4321 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4381 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4441 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4501 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4561 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4621 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4681 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4741 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4801 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4861 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4921 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4981 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5041 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5101 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5161 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5221 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5281 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5341 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5401 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5461 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6301 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6361 cgccatgcat
//
