LOCUS       sequence-355449-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP sensor of Kras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  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 6386)
  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-Kras
FEATURES             Location/Qualifiers
     source          1..6386
                     /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             2418..2984
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     misc_feature    2985..3050
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3174..3295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3211..3230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3265..3284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3302..3757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3439..3460)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3651..3670
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3784..3888
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3890..4247
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3890..4081
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3913..3933)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4098..4233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4282..5076
                     /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             4282..5076
                     /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(4336..4355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4946..4965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5264..5283)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5308..5355
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5684..6272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6173..6192
                     /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 gatctagcct gctgaaaatg actgaatata aacttgtggt agttggagct ggtggcgtag
     2461 gcaagagtgc cttgacgata cagctaattc agaatcattt tgtggacgaa tatgatccaa
     2521 caatagagga ttcctacagg aagcaagtag taattgatgg agaaacctgt ctcttggata
     2581 ttctcgacac agcaggtcaa gaggagtaca gtgcaatgag ggaccagtac atgaggactg
     2641 gggagggctt tctttgtgta tttgccataa ataatactaa atcatttgaa gatattcacc
     2701 attatagaga acaaattaaa agagttaagg actctgaaga tgtacctatg gtcctagtag
     2761 gaaataaatg tgatttgcct tctagaacag tagacacaaa acaggctcag gacttagcaa
     2821 gaagttatgg aattcctttt attgaaacat cagcaaagac aagacagggt gttgatgatg
     2881 ccttctatac attagttcga gaaattcgaa aacataaaga aaagatgagc aaagatggta
     2941 aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtaatccgga ctcagatctc
     3001 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     3061 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3121 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3181 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3241 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     3301 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3361 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3421 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3481 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3541 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3601 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3661 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3721 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3901 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4081 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4141 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4201 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4261 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4321 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4381 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4441 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4501 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4561 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4621 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4681 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4741 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4801 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4861 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4921 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4981 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5041 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5101 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5161 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5221 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5281 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5341 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5401 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5461 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5521 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5581 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5641 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5701 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5761 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5821 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5881 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5941 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6001 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6061 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6301 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6361 tggataaccg tattaccgcc atgcat
//
