LOCUS       sequence-468953-        8780 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of the R487L mutant variant of human PDGFRA, which might 
            be associated with Gastrointestinal Stromal Tumor.
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 Apr 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-R487L-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        327..530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     575..594
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        575..593
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3990..4031
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4041..4058
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4081..4098)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4087..4311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4357..4785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4444..4463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4654..4675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4794..4814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4799..5128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4979..5114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5041..5060
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5195..5989
                     /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(5249..5268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5859..5878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6165..6286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6202..6221)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6256..6275
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6335..6351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6335..6351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6348..6370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6359..6375
                     /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(6383..6389,6390..6407,6408..6413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6428..6449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6566..6583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6737..7325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6817..6836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7496..8287,8288..8356))
                     /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     8119..8138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8357..8461)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8536..8555)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8727..8780,1..326)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
       61 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      301 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      361 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      421 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      481 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      541 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      601 aagctggcta gttaagcttg gtaccgagct cggatccagt acccttcacc atggggactt
      661 cccatccggc gttcctggtc ttaggctgtc ttctcacagg gctgagccta atcctctgcc
      721 agctttcatt accctctatc cttccaaatg aaaatgaaaa ggttgtgcag ctgaattcat
      781 ccttttctct gagatgcttt ggggagagtg aagtgagctg gcagtacccc atgtctgaag
      841 aagagagctc cgatgtggaa atcagaaatg aagaaaacaa cagcggcctt tttgtgacgg
      901 tcttggaagt gagcagtgcc tcggcggccc acacagggtt gtacacttgc tattacaacc
      961 acactcagac agaagagaat gagcttgaag gcaggcacat ttacatctat gtgccagacc
     1021 cagatgtagc ctttgtacct ctaggaatga cggattattt agtcatcgtg gaggatgatg
     1081 attctgccat tataccttgt cgcacaactg atcccgagac tcctgtaacc ttacacaaca
     1141 gtgagggggt ggtacctgcc tcctacgaca gcagacaggg ctttaatggg accttcactg
     1201 tagggcccta tatctgtgag gccaccgtca aaggaaagaa gttccagacc atcccattta
     1261 atgtttatgc tttaaaagca acatcagagc tggatctaga aatggaagct cttaaaaccg
     1321 tgtataagtc aggggaaacg attgtggtca cctgtgctgt ttttaacaat gaggtggttg
     1381 accttcaatg gacttaccct ggagaagtga aaggcaaagg catcacaatg ctggaagaaa
     1441 tcaaagtccc atccatcaaa ttggtgtaca ctttgacggt ccccgaggcc acggtgaaag
     1501 acagtggaga ttacgaatgt gctgcccgcc aggctaccag ggaggtcaaa gaaatgaaga
     1561 aagtcactat ttctgtccat gagaaaggtt tcattgaaat caaacccacc ttcagccagt
     1621 tggaagctgt caacctgcat gaagtcaaac attttgttgt agaggtgcgg gcctacccac
     1681 ctcccaggat atcctggctg aaaaacaatc tgactctgat tgaaaatctc actgagatca
     1741 ccactgatgt ggaaaagatt caggaaataa ggtatcgaag caaattaaag ctgatccgtg
     1801 ctaaggaaga agacagtggc cattatacta ttgtagctca aaatgaagat gctgtgaaga
     1861 gctatacttt tgaactgtta actcaagttc cttcatccat tctggacttg gtcgatgatc
     1921 accatggctc aactggggga cagacggtga ggtgcacagc tgaaggcacg ccgcttcctg
     1981 atattgagtg gatgatatgc aaagatatta agaaatgtaa taatgaaact tcctggacta
     2041 ttttggccaa caatgtctca aacatcatca cggagatcca ctcccgagac aggagtaccg
     2101 tggagggcct tgtgactttc gccaaagtgg aggagaccat cgccgtgcga tgcctggcta
     2161 agaatctcct tggagctgag aaccgagagc tgaagctggt ggctcccacc ctgcgttctg
     2221 aactcacggt ggctgctgca gtcctggtgc tgttggtgat tgtgatcatc tcacttattg
     2281 tcctggttgt catttggaaa cagaaaccga ggtatgaaat tcgctggagg gtcattgaat
     2341 caatcagccc ggatggacat gaatatattt atgtggaccc gatgcagctg ccttatgact
     2401 caagatggga gtttccaaga gatggactag tgcttggtcg ggtcttgggg tctggagcgt
     2461 ttgggaaggt ggttgaagga acagcctatg gattaagccg gtcccaacct gtcatgaaag
     2521 ttgcagtgaa gatgctaaaa cccacggcca gatccagtga aaaacaagct ctcatgtctg
     2581 aactgaagat aatgactcac ctggggccac atttgaacat tgtaaacttg ctgggagcct
     2641 gcaccaagtc aggccccatt tacatcatca cagagtattg cttctatgga gatttggtca
     2701 actatttgca taagaatagg gatagcttcc tgagccacca cccagagaag ccaaagaaag
     2761 agctggatat ctttggattg aaccctgctg atgaaagcac acggagctat gttattttat
     2821 cttttgaaaa caatggtgac tacatggaca tgaagcaggc tgatactaca cagtatgtcc
     2881 ccatgctaga aaggaaagag gtttctaaat attccgacat ccagagatca ctctatgatc
     2941 gtccagcctc atataagaag aaatctatgt tagactcaga agtcaaaaac ctcctttcag
     3001 atgataactc agaaggcctt actttattgg atttgttgag cttcacctat caagttgccc
     3061 gaggaatgga gtttttggct tcaaaaaatt gtgtccaccg tgatctggct gctcgcaacg
     3121 tcctcctggc acaaggaaaa attgtgaaga tctgtgactt tggcctggcc agagacatca
     3181 tgcatgattc gaactatgtg tcgaaaggca gtacctttct gcccgtgaag tggatggctc
     3241 ctgagagcat ctttgacaac ctctacacca cactgagtga tgtctggtct tatggcattc
     3301 tgctctggga gatcttttcc cttggtggca ccccttaccc cggcatgatg gtggattcta
     3361 ctttctacaa taagatcaag agtgggtacc ggatggccaa gcctgaccac gctaccagtg
     3421 aagtctacga gatcatggtg aaatgctgga acagtgagcc ggagaagaga ccctcctttt
     3481 accacctgag tgagattgtg gagaatctgc tgcctggaca atataaaaag agttatgaaa
     3541 aaattcacct ggacttcctg aagagtgacc atcctgctgt ggcacgcatg cgtgtggact
     3601 cagacaatgc atacattggt gtcacctaca aaaacgagga agacaagctg aaggactggg
     3661 agggtggtct ggatgagcag agactgagcg ctgacagtgg ctacatcatt cctctgcctg
     3721 acattgaccc tgtccctgag gaggaggacc tgggcaagag gaacagacac agctcgcaga
     3781 cctctgaaga gagtgccatt gagacgggtt ccagcagttc caccttcatc aagagagagg
     3841 acgagaccat tgaagacatc gacatgatgg acgacatcgg catagactct tcagacctgg
     3901 tggaagacag ctttcttaag ggtcaagaca attctgcaga tatccagcac agtggcggcc
     3961 gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     4021 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     4081 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4141 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4201 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4261 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4321 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4381 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4441 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4501 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4561 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4621 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4681 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4741 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4801 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4861 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4921 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4981 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5041 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5101 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5161 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5221 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5281 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5341 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5401 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5461 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5521 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5581 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5641 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5701 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5761 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5821 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5881 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5941 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6001 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6061 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6121 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6301 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6481 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6541 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6601 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7501 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7561 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7621 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7681 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7741 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7801 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7861 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7921 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7981 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8041 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8101 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8161 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8221 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8281 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8341 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8401 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8461 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     8521 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     8581 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     8641 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     8701 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     8761 aattacgggg tcattagttc
//
