LOCUS       sequence-355232-        6407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddRFP sensor of Kras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  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 6407)
  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-R-Kras
FEATURES             Location/Qualifiers
     source          1..6407
                     /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             2439..3005
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     misc_feature    3006..3071
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3195..3316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3232..3251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3286..3305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3323..3778)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3460..3481)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3672..3691
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3805..3909
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3911..4268
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3911..4102
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3934..3954)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4119..4254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4303..5097
                     /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             4303..5097
                     /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(4357..4376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4967..4986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5285..5304)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5329..5376
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5705..6293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6194..6213
                     /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 gggccgcccc tacgagggca
     1801 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     1861 tgtcccccca gatcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacgtccccg
     1921 attacatgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg cacttcgagg
     1981 acggcggtct ggtgaccgtg actcaggaca cctccctgca ggacggcacg ctgatctaca
     2041 aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacct
     2101 tgggctggga ttatgccacc gagcgcctgt accccgaaga cggcgtgctg aagggcgagc
     2161 ttctggggcg cctgaagctg aaggacggcg gcctcaacct ggtggagttc aagaccatct
     2221 acatggccaa gaagcccgtg caactgcccg gctactactt cgtggacacc aagctgggca
     2281 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc
     2341 accacctggg gcatggcacc ggcagcaccg gcagcggcag ctccggtacc gcctcctccg
     2401 aggacaacaa cctgtacaag agatctagcc tgctgaaaat gactgaatat aaacttgtgg
     2461 tagttggagc tggtggcgta ggcaagagtg ccttgacgat acagctaatt cagaatcatt
     2521 ttgtggacga atatgatcca acaatagagg attcctacag gaagcaagta gtaattgatg
     2581 gagaaacctg tctcttggat attctcgaca cagcaggtca agaggagtac agtgcaatga
     2641 gggaccagta catgaggact ggggagggct ttctttgtgt atttgccata aataatacta
     2701 aatcatttga agatattcac cattatagag aacaaattaa aagagttaag gactctgaag
     2761 atgtacctat ggtcctagta ggaaataaat gtgatttgcc ttctagaaca gtagacacaa
     2821 aacaggctca ggacttagca agaagttatg gaattccttt tattgaaaca tcagcaaaga
     2881 caagacaggg tgttgatgat gccttctata cattagttcg agaaattcga aaacataaag
     2941 aaaagatgag caaagatggt aaaaagaaga aaaagaagtc aaagacaaag tgtgtaatta
     3001 tgtaatccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     3061 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     3121 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3181 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3241 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3301 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3361 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3421 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3481 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3541 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3601 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3661 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3721 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3781 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3841 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3901 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4141 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4201 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4261 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4321 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4381 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4441 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4501 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4561 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4621 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4681 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4741 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4801 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4861 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4921 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4981 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5041 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5101 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5161 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5221 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5281 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5341 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5401 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5461 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5521 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5581 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5641 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5821 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6061 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6301 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6361 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
