LOCUS       sequence-468865-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the T674I mutant variant of human PDGFRA, which is 
            associated with Idiopathic Hypereosinophilic Syndrome, resistant to 
            imatinib .
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-T674I-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1578..1619
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1629..1646
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1669..1686)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1675..1899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1945..2373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2032..2051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2242..2263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2382..2402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2387..2716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2567..2702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2629..2648
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2783..3577
                     /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(2837..2856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3447..3466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3753..3874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3790..3809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3844..3863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3923..3939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3923..3939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3936..3958)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3947..3963
                     /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(3971..3977,3978..3995,3996..4001))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4016..4037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4154..4171)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4325..4913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4405..4424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5084..5875,5876..5944))
                     /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     5707..5726
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5945..6049)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6124..6143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6315..6694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6695..6898
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6849..6869
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6943..6962
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6943..6961
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttccaagaga tggactagtg cttggtcggg tcttggggtc tggagcgttt gggaaggtgg
       61 ttgaaggaac agcctatgga ttaagccggt cccaacctgt catgaaagtt gcagtgaaga
      121 tgctaaaacc cacggccaga tccagtgaaa aacaagctct catgtctgaa ctgaagataa
      181 tgactcacct ggggccacat ttgaacattg taaacttgct gggagcctgc accaagtcag
      241 gccccattta catcatcata gagtattgct tctatggaga tttggtcaac tatttgcata
      301 agaataggga tagcttcctg agccaccacc cagagaagcc aaagaaagag ctggatatct
      361 ttggattgaa ccctgctgat gaaagcacac ggagctatgt tattttatct tttgaaaaca
      421 atggtgacta catggacatg aagcaggctg atactacaca gtatgtcccc atgctagaaa
      481 ggaaagaggt ttctaaatat tccgacatcc agagatcact ctatgatcgt ccagcctcat
      541 ataagaagaa atctatgtta gactcagaag tcaaaaacct cctttcagat gataactcag
      601 aaggccttac tttattggat ttgttgagct tcacctatca agttgcccga ggaatggagt
      661 ttttggcttc aaaaaattgt gtccaccgtg atctggctgc tcgcaacgtc ctcctggcac
      721 aaggaaaaat tgtgaagatc tgtgactttg gcctggccag agacatcatg catgattcga
      781 actatgtgtc gaaaggcagt acctttctgc ccgtgaagtg gatggctcct gagagcatct
      841 ttgacaacct ctacaccaca ctgagtgatg tctggtctta tggcattctg ctctgggaga
      901 tcttttccct tggtggcacc ccttaccccg gcatgatggt ggattctact ttctacaata
      961 agatcaagag tgggtaccgg atggccaagc ctgaccacgc taccagtgaa gtctacgaga
     1021 tcatggtgaa atgctggaac agtgagccgg agaagagacc ctccttttac cacctgagtg
     1081 agattgtgga gaatctgctg cctggacaat ataaaaagag ttatgaaaaa attcacctgg
     1141 acttcctgaa gagtgaccat cctgctgtgg cacgcatgcg tgtggactca gacaatgcat
     1201 acattggtgt cacctacaaa aacgaggaag acaagctgaa ggactgggag ggtggtctgg
     1261 atgagcagag actgagcgct gacagtggct acatcattcc tctgcctgac attgaccctg
     1321 tccctgagga ggaggacctg ggcaagagga acagacacag ctcgcagacc tctgaagaga
     1381 gtgccattga gacgggttcc agcagttcca ccttcatcaa gagagaggac gagaccattg
     1441 aagacatcga catgatggac gacatcggca tagactcttc agacctggtg gaagacagct
     1501 ttcttaaggg tcaagacaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     1561 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     1621 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     1681 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1741 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1801 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1861 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     1921 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2101 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2161 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2281 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2341 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2461 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2521 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2581 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2641 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2701 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     2761 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2821 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2881 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2941 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3001 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3061 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3121 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3181 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3241 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3301 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3361 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3421 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3481 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3541 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     3601 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3661 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3721 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3781 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3841 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3901 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4741 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5041 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5101 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5161 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5221 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5281 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5341 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5401 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5461 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5521 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5581 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5641 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5701 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5761 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5821 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5881 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5941 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6001 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6061 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     6121 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     6181 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     6241 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     6301 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     6361 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6421 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6481 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     6541 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6601 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6661 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     6721 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     6781 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6841 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     6901 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     6961 ggagacccaa gctggctagt taagcttggt accgagctcg gatccagtac ccttcaccat
     7021 ggggacttcc catccggcgt tcctggtctt aggctgtctt ctcacagggc tgagcctaat
     7081 cctctgccag ctttcattac cctctatcct tccaaatgaa aatgaaaagg ttgtgcagct
     7141 gaattcatcc ttttctctga gatgctttgg ggagagtgaa gtgagctggc agtaccccat
     7201 gtctgaagaa gagagctccg atgtggaaat cagaaatgaa gaaaacaaca gcggcctttt
     7261 tgtgacggtc ttggaagtga gcagtgcctc ggcggcccac acagggttgt acacttgcta
     7321 ttacaaccac actcagacag aagagaatga gcttgaaggc aggcacattt acatctatgt
     7381 gccagaccca gatgtagcct ttgtacctct aggaatgacg gattatttag tcatcgtgga
     7441 ggatgatgat tctgccatta taccttgtcg cacaactgat cccgagactc ctgtaacctt
     7501 acacaacagt gagggggtgg tacctgcctc ctacgacagc agacagggct ttaatgggac
     7561 cttcactgta gggccctata tctgtgaggc caccgtcaaa ggaaagaagt tccagaccat
     7621 cccatttaat gtttatgctt taaaagcaac atcagagctg gatctagaaa tggaagctct
     7681 taaaaccgtg tataagtcag gggaaacgat tgtggtcacc tgtgctgttt ttaacaatga
     7741 ggtggttgac cttcaatgga cttaccctgg agaagtgaaa ggcaaaggca tcacaatgct
     7801 ggaagaaatc aaagtcccat ccatcaaatt ggtgtacact ttgacggtcc ccgaggccac
     7861 ggtgaaagac agtggagatt acgaatgtgc tgcccgccag gctaccaggg aggtcaaaga
     7921 aatgaagaaa gtcactattt ctgtccatga gaaaggtttc attgaaatca aacccacctt
     7981 cagccagttg gaagctgtca acctgcatga agtcaaacat tttgttgtag aggtgcgggc
     8041 ctacccacct cccaggatat cctggctgaa aaacaatctg actctgattg aaaatctcac
     8101 tgagatcacc actgatgtgg aaaagattca ggaaataagg tatcgaagca aattaaagct
     8161 gatccgtgct aaggaagaag acagtggcca ttatactatt gtagctcaaa atgaagatgc
     8221 tgtgaagagc tatacttttg aactgttaac tcaagttcct tcatccattc tggacttggt
     8281 cgatgatcac catggctcaa ctgggggaca gacggtgagg tgcacagctg aaggcacgcc
     8341 gcttcctgat attgagtgga tgatatgcaa agatattaag aaatgtaata atgaaacttc
     8401 ctggactatt ttggccaaca atgtctcaaa catcatcacg gagatccact cccgagacag
     8461 gagtaccgtg gagggccgtg tgactttcgc caaagtggag gagaccatcg ccgtgcgatg
     8521 cctggctaag aatctccttg gagctgagaa ccgagagctg aagctggtgg ctcccaccct
     8581 gcgttctgaa ctcacggtgg ctgctgcagt cctggtgctg ttggtgattg tgatcatctc
     8641 acttattgtc ctggttgtca tttggaaaca gaaaccgagg tatgaaattc gctggagggt
     8701 cattgaatca atcagcccgg atggacatga atatatttat gtggacccga tgcagctgcc
     8761 ttatgactca agatgggagt
//
