LOCUS       sequence-468956-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the V561D mutant variant of human PDGFRA, which is 
            associated with Gastrointestinal Stromal Tumor and GIST-Plus 
            Syndrome, , constantly activated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8780)
  AUTHORS   Gudernova I, Balek L, Varecha M, Kucerova JF, Kunova Bosakova M, 
            Fafilek B, Palusova V, Uldrijan S, Trantirek L, Krejci P
  TITLE     Inhibitor repurposing reveals ALK, LTK, FGFR, RET and TRK kinases as
            the targets of AZD1480.
  JOURNAL   Oncotarget. 2017 Nov 27;8(65):109319-109331. doi: 
            10.18632/oncotarget.22674. eCollection 2017 Dec 12.
  PUBMED    29312610
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"; doi: 
            "10.18632/oncotarget.22674"; journalName: "Oncotarget"; date: 
            "2017-11-27- 27"; volume: "8"; issue: "65"; pages: "109319-109331"
COMMENT     Sequence Label: pcDNA3.1-PDGFRA-V561D-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..65
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             200..994
                     /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(254..273)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     864..883
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1170..1291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1207..1226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1261..1280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1340..1356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1340..1356)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1353..1375)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1364..1380
                     /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(1388..1394,1395..1412,1413..1418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1433..1454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1571..1588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1742..2330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1822..1841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2501..3292,3293..3361))
                     /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     3124..3143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3362..3466)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3541..3560)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3732..4111
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4112..4315
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4266..4286
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4360..4379
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4360..4378
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7775..7816
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7826..7843
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7866..7883)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7872..8096
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8142..8570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8229..8248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8439..8460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8579..8599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8584..8780,1..133)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8764..8780,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
       61 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
      121 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
      181 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
      241 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
      301 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
      361 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
      421 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
      481 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
      541 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
      601 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
      661 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
      721 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
      781 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
      841 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
      901 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
      961 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     1021 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     1081 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     1141 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     1201 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1261 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     1321 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1381 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1441 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1501 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1561 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1621 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1681 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1741 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1801 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1861 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1921 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1981 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2041 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2101 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2161 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2221 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2401 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2461 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2521 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2581 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2641 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2701 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2761 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2821 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2881 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2941 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3001 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3061 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3121 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3181 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3241 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3301 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3361 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3421 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3481 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     3541 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     3601 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     3661 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     3721 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     3781 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3841 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3901 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3961 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4021 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4081 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4141 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4201 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4261 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     4321 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     4381 gacccaagct ggctagttaa gcttggtacc gagctcggat ccagtaccct tcaccatggg
     4441 gacttcccat ccggcgttcc tggtcttagg ctgtcttctc acagggctga gcctaatcct
     4501 ctgccagctt tcattaccct ctatccttcc aaatgaaaat gaaaaggttg tgcagctgaa
     4561 ttcatccttt tctctgagat gctttgggga gagtgaagtg agctggcagt accccatgtc
     4621 tgaagaagag agctccgatg tggaaatcag aaatgaagaa aacaacagcg gcctttttgt
     4681 gacggtcttg gaagtgagca gtgcctcggc ggcccacaca gggttgtaca cttgctatta
     4741 caaccacact cagacagaag agaatgagct tgaaggcagg cacatttaca tctatgtgcc
     4801 agacccagat gtagcctttg tacctctagg aatgacggat tatttagtca tcgtggagga
     4861 tgatgattct gccattatac cttgtcgcac aactgatccc gagactcctg taaccttaca
     4921 caacagtgag ggggtggtac ctgcctccta cgacagcaga cagggcttta atgggacctt
     4981 cactgtaggg ccctatatct gtgaggccac cgtcaaagga aagaagttcc agaccatccc
     5041 atttaatgtt tatgctttaa aagcaacatc agagctggat ctagaaatgg aagctcttaa
     5101 aaccgtgtat aagtcagggg aaacgattgt ggtcacctgt gctgttttta acaatgaggt
     5161 ggttgacctt caatggactt accctggaga agtgaaaggc aaaggcatca caatgctgga
     5221 agaaatcaaa gtcccatcca tcaaattggt gtacactttg acggtccccg aggccacggt
     5281 gaaagacagt ggagattacg aatgtgctgc ccgccaggct accagggagg tcaaagaaat
     5341 gaagaaagtc actatttctg tccatgagaa aggtttcatt gaaatcaaac ccaccttcag
     5401 ccagttggaa gctgtcaacc tgcatgaagt caaacatttt gttgtagagg tgcgggccta
     5461 cccacctccc aggatatcct ggctgaaaaa caatctgact ctgattgaaa atctcactga
     5521 gatcaccact gatgtggaaa agattcagga aataaggtat cgaagcaaat taaagctgat
     5581 ccgtgctaag gaagaagaca gtggccatta tactattgta gctcaaaatg aagatgctgt
     5641 gaagagctat acttttgaac tgttaactca agttccttca tccattctgg acttggtcga
     5701 tgatcaccat ggctcaactg ggggacagac ggtgaggtgc acagctgaag gcacgccgct
     5761 tcctgatatt gagtggatga tatgcaaaga tattaagaaa tgtaataatg aaacttcctg
     5821 gactattttg gccaacaatg tctcaaacat catcacggag atccactccc gagacaggag
     5881 taccgtggag ggccgtgtga ctttcgccaa agtggaggag accatcgccg tgcgatgcct
     5941 ggctaagaat ctccttggag ctgagaaccg agagctgaag ctggtggctc ccaccctgcg
     6001 ttctgaactc acggtggctg ctgcagtcct ggtgctgttg gtgattgtga tcatctcact
     6061 tattgtcctg gttgtcattt ggaaacagaa accgaggtat gaaattcgct ggagggacat
     6121 tgaatcaatc agcccggatg gacatgaata tatttatgtg gacccgatgc agctgcctta
     6181 tgactcaaga tgggagtttc caagagatgg actagtgctt ggtcgggtct tggggtctgg
     6241 agcgtttggg aaggtggttg aaggaacagc ctatggatta agccggtccc aacctgtcat
     6301 gaaagttgca gtgaagatgc taaaacccac ggccagatcc agtgaaaaac aagctctcat
     6361 gtctgaactg aagataatga ctcacctggg gccacatttg aacattgtaa acttgctggg
     6421 agcctgcacc aagtcaggcc ccatttacat catcacagag tattgcttct atggagattt
     6481 ggtcaactat ttgcataaga atagggatag cttcctgagc caccacccag agaagccaaa
     6541 gaaagagctg gatatctttg gattgaaccc tgctgatgaa agcacacgga gctatgttat
     6601 tttatctttt gaaaacaatg gtgactacat ggacatgaag caggctgata ctacacagta
     6661 tgtccccatg ctagaaagga aagaggtttc taaatattcc gacatccaga gatcactcta
     6721 tgatcgtcca gcctcatata agaagaaatc tatgttagac tcagaagtca aaaacctcct
     6781 ttcagatgat aactcagaag gccttacttt attggatttg ttgagcttca cctatcaagt
     6841 tgcccgagga atggagtttt tggcttcaaa aaattgtgtc caccgtgatc tggctgctcg
     6901 caacgtcctc ctggcacaag gaaaaattgt gaagatctgt gactttggcc tggccagaga
     6961 catcatgcat gattcgaact atgtgtcgaa aggcagtacc tttctgcccg tgaagtggat
     7021 ggctcctgag agcatctttg acaacctcta caccacactg agtgatgtct ggtcttatgg
     7081 cattctgctc tgggagatct tttcccttgg tggcacccct taccccggca tgatggtgga
     7141 ttctactttc tacaataaga tcaagagtgg gtaccggatg gccaagcctg accacgctac
     7201 cagtgaagtc tacgagatca tggtgaaatg ctggaacagt gagccggaga agagaccctc
     7261 cttttaccac ctgagtgaga ttgtggagaa tctgctgcct ggacaatata aaaagagtta
     7321 tgaaaaaatt cacctggact tcctgaagag tgaccatcct gctgtggcac gcatgcgtgt
     7381 ggactcagac aatgcataca ttggtgtcac ctacaaaaac gaggaagaca agctgaagga
     7441 ctgggagggt ggtctggatg agcagagact gagcgctgac agtggctaca tcattcctct
     7501 gcctgacatt gaccctgtcc ctgaggagga ggacctgggc aagaggaaca gacacagctc
     7561 gcagacctct gaagagagtg ccattgagac gggttccagc agttccacct tcatcaagag
     7621 agaggacgag accattgaag acatcgacat gatggacgac atcggcatag actcttcaga
     7681 cctggtggaa gacagctttc ttaagggtca agacaattct gcagatatcc agcacagtgg
     7741 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     7801 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     7861 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     7921 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     7981 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     8041 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     8101 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     8161 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     8221 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     8281 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     8341 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     8401 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     8461 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     8521 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     8581 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     8641 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     8701 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     8761 cccatcccgc ccctaactcc
//
