LOCUS       sequence-466512-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the K627M kinase defective mutant variant of human 
            PDGFRA .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8780)
  AUTHORS   Gudernova I, Foldynova-Trantirkova S, Ghannamova BE, Fafilek B, 
            Varecha M, Balek L, Hruba E, Jonatova L, Jelinkova I, Kunova 
            Bosakova M, Trantirek L, Mayer J, Krejci P
  TITLE     One reporter for in-cell activity profiling of majority of protein 
            kinase oncogenes.
  JOURNAL   Elife. 2017 Feb 15;6. pii: e21536. doi: 10.7554/eLife.21536.
  PUBMED    28199182
REFERENCE   2  (bases 1 to 8780)
  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: "Elife."; 
            date: "2017-02-15"; volume: "6. pii: e21536. doi"; pages: " 
            10.7554/eLife.21536"
COMMENT     Sequence Label: pcDNA3.1-PDGFRA-K627M-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4278..4319
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4329..4346
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4369..4386)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4375..4599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4645..5073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4732..4751)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4942..4963
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5082..5102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5087..5416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5267..5402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5329..5348
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5483..6277
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5537..5556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6147..6166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6453..6574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6490..6509)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6544..6563
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6623..6639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6623..6639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6636..6658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6647..6663
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6671..6677,6678..6695,6696..6701))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6716..6737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6854..6871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7025..7613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7105..7124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7784..8575,8576..8644))
                     /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     8407..8426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8645..8749)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccagtac ccttcaccat ggggacttcc catccggcgt
      961 tcctggtctt aggctgtctt ctcacagggc tgagcctaat cctctgccag ctttcattac
     1021 cctctatcct tccaaatgaa aatgaaaagg ttgtgcagct gaattcatcc ttttctctga
     1081 gatgctttgg ggagagtgaa gtgagctggc agtaccccat gtctgaagaa gagagctccg
     1141 atgtggaaat cagaaatgaa gaaaacaaca gcggcctttt tgtgacggtc ttggaagtga
     1201 gcagtgcctc ggcggcccac acagggttgt acacttgcta ttacaaccac actcagacag
     1261 aagagaatga gcttgaaggc aggcacattt acatctatgt gccagaccca gatgtagcct
     1321 ttgtacctct aggaatgacg gattatttag tcatcgtgga ggatgatgat tctgccatta
     1381 taccttgtcg cacaactgat cccgagactc ctgtaacctt acacaacagt gagggggtgg
     1441 tacctgcctc ctacgacagc agacagggct ttaatgggac cttcactgta gggccctata
     1501 tctgtgaggc caccgtcaaa ggaaagaagt tccagaccat cccatttaat gtttatgctt
     1561 taaaagcaac atcagagctg gatctagaaa tggaagctct taaaaccgtg tataagtcag
     1621 gggaaacgat tgtggtcacc tgtgctgttt ttaacaatga ggtggttgac cttcaatgga
     1681 cttaccctgg agaagtgaaa ggcaaaggca tcacaatgct ggaagaaatc aaagtcccat
     1741 ccatcaaatt ggtgtacact ttgacggtcc ccgaggccac ggtgaaagac agtggagatt
     1801 acgaatgtgc tgcccgccag gctaccaggg aggtcaaaga aatgaagaaa gtcactattt
     1861 ctgtccatga gaaaggtttc attgaaatca aacccacctt cagccagttg gaagctgtca
     1921 acctgcatga agtcaaacat tttgttgtag aggtgcgggc ctacccacct cccaggatat
     1981 cctggctgaa aaacaatctg actctgattg aaaatctcac tgagatcacc actgatgtgg
     2041 aaaagattca ggaaataagg tatcgaagca aattaaagct gatccgtgct aaggaagaag
     2101 acagtggcca ttatactatt gtagctcaaa atgaagatgc tgtgaagagc tatacttttg
     2161 aactgttaac tcaagttcct tcatccattc tggacttggt cgatgatcac catggctcaa
     2221 ctgggggaca gacggtgagg tgcacagctg aaggcacgcc gcttcctgat attgagtgga
     2281 tgatatgcaa agatattaag aaatgtaata atgaaacttc ctggactatt ttggccaaca
     2341 atgtctcaaa catcatcacg gagatccact cccgagacag gagtaccgtg gagggccgtg
     2401 tgactttcgc caaagtggag gagaccatcg ccgtgcgatg cctggctaag aatctccttg
     2461 gagctgagaa ccgagagctg aagctggtgg ctcccaccct gcgttctgaa ctcacggtgg
     2521 ctgctgcagt cctggtgctg ttggtgattg tgatcatctc acttattgtc ctggttgtca
     2581 tttggaaaca gaaaccgagg tatgaaattc gctggagggt cattgaatca atcagcccgg
     2641 atggacatga atatatttat gtggacccga tgcagctgcc ttatgactca agatgggagt
     2701 ttccaagaga tggactagtg cttggtcggg tcttggggtc tggagcgttt gggaaggtgg
     2761 ttgaaggaac agcctatgga ttaagccggt cccaacctgt catgaaagtt gcagtgatga
     2821 tgctaaaacc cacggccaga tccagtgaaa aacaagctct catgtctgaa ctgaagataa
     2881 tgactcacct ggggccacat ttgaacattg taaacttgct gggagcctgc accaagtcag
     2941 gccccattta catcatcaca gagtattgct tctatggaga tttggtcaac tatttgcata
     3001 agaataggga tagcttcctg agccaccacc cagagaagcc aaagaaagag ctggatatct
     3061 ttggattgaa ccctgctgat gaaagcacac ggagctatgt tattttatct tttgaaaaca
     3121 atggtgacta catggacatg aagcaggctg atactacaca gtatgtcccc atgctagaaa
     3181 ggaaagaggt ttctaaatat tccgacatcc agagatcact ctatgatcgt ccagcctcat
     3241 ataagaagaa atctatgtta gactcagaag tcaaaaacct cctttcagat gataactcag
     3301 aaggccttac tttattggat ttgttgagct tcacctatca agttgcccga ggaatggagt
     3361 ttttggcttc aaaaaattgt gtccaccgtg atctggctgc tcgcaacgtc ctcctggcac
     3421 aaggaaaaat tgtgaagatc tgtgactttg gcctggccag agacatcatg catgattcga
     3481 actatgtgtc gaaaggcagt acctttctgc ccgtgaagtg gatggctcct gagagcatct
     3541 ttgacaacct ctacaccaca ctgagtgatg tctggtctta tggcattctg ctctgggaga
     3601 tcttttccct tggtggcacc ccttaccccg gcatgatggt ggattctact ttctacaata
     3661 agatcaagag tgggtaccgg atggccaagc ctgaccacgc taccagtgaa gtctacgaga
     3721 tcatggtgaa atgctggaac agtgagccgg agaagagacc ctccttttac cacctgagtg
     3781 agattgtgga gaatctgctg cctggacaat ataaaaagag ttatgaaaaa attcacctgg
     3841 acttcctgaa gagtgaccat cctgctgtgg cacgcatgcg tgtggactca gacaatgcat
     3901 acattggtgt cacctacaaa aacgaggaag acaagctgaa ggactgggag ggtggtctgg
     3961 atgagcagag actgagcgct gacagtggct acatcattcc tctgcctgac attgaccctg
     4021 tccctgagga ggaggacctg ggcaagagga acagacacag ctcgcagacc tctgaagaga
     4081 gtgccattga gacgggttcc agcagttcca ccttcatcaa gagagaggac gagaccattg
     4141 aagacatcga catgatggac gacatcggca tagactcttc agacctggtg gaagacagct
     4201 ttcttaaggg tcaagacaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     4261 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     4321 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     4381 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4441 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4501 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4561 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4621 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4681 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4741 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4801 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4861 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4921 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4981 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5041 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     5101 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5161 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5221 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5281 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5341 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5401 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     5461 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5521 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5581 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5641 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5701 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5761 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5821 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5881 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5941 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6001 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6061 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6121 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6181 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6241 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     6301 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6361 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6421 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     6481 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6541 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6601 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6661 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6721 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6781 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6841 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6901 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6961 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7441 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7741 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7801 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7861 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7921 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7981 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8041 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8101 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8161 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8221 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8281 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8341 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8401 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8461 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8521 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8581 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8641 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8701 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8761 gaaaagtgcc acctgacgtc
//
