LOCUS       sequence-468951-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the G853D mutant variant of human PDGFRA, which might 
            be associated with Melanoma .
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-G853D-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1756..1797
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1807..1824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1847..1864)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1853..2077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2123..2551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2210..2229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2420..2441
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2560..2580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2565..2894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2745..2880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2807..2826
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2961..3755
                     /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(3015..3034)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3625..3644
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3931..4052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3968..3987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4022..4041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4101..4117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4101..4117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4114..4136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4125..4141
                     /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(4149..4155,4156..4173,4174..4179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4194..4215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4332..4349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4503..5091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4583..4602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5262..6053,6054..6122))
                     /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     5885..5904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6123..6227)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6302..6321)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6493..6872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6873..7076
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7027..7047
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7121..7140
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7121..7139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctgcagtcc tggtgctgtt ggtgattgtg atcatctcac ttattgtcct ggttgtcatt
       61 tggaaacaga aaccgaggta tgaaattcgc tggagggtca ttgaatcaat cagcccggat
      121 ggacatgaat atatttatgt ggacccgatg cagctgcctt atgactcaag atgggagttt
      181 ccaagagatg gactagtgct tggtcgggtc ttggggtctg gagcgtttgg gaaggtggtt
      241 gaaggaacag cctatggatt aagccggtcc caacctgtca tgaaagttgc agtgaagatg
      301 ctaaaaccca cggccagatc cagtgaaaaa caagctctca tgtctgaact gaagataatg
      361 actcacctgg ggccacattt gaacattgta aacttgctgg gagcctgcac caagtcaggc
      421 cccatttaca tcatcacaga gtattgcttc tatggagatt tggtcaacta tttgcataag
      481 aatagggata gcttcctgag ccaccaccca gagaagccaa agaaagagct ggatatcttt
      541 ggattgaacc ctgctgatga aagcacacgg agctatgtta ttttatcttt tgaaaacaat
      601 ggtgactaca tggacatgaa gcaggctgat actacacagt atgtccccat gctagaaagg
      661 aaagaggttt ctaaatattc cgacatccag agatcactct atgatcgtcc agcctcatat
      721 aagaagaaat ctatgttaga ctcagaagtc aaaaacctcc tttcagatga taactcagaa
      781 ggccttactt tattggattt gttgagcttc acctatcaag ttgcccgagg aatggagttt
      841 ttggcttcaa aaaattgtgt ccaccgtgat ctggctgctc gcaacgtcct cctggcacaa
      901 ggaaaaattg tgaagatctg tgactttggc ctggccagag acatcatgca tgattcgaac
      961 tatgtgtcga aagacagtac ctttctgccc gtgaagtgga tggctcctga gagcatcttt
     1021 gacaacctct acaccacact gagtgatgtc tggtcttatg gcattctgct ctgggagatc
     1081 ttttcccttg gtggcacccc ttaccccggc atgatggtgg attctacttt ctacaataag
     1141 atcaagagtg ggtaccggat ggccaagcct gaccacgcta ccagtgaagt ctacgagatc
     1201 atggtgaaat gctggaacag tgagccggag aagagaccct ccttttacca cctgagtgag
     1261 attgtggaga atctgctgcc tggacaatat aaaaagagtt atgaaaaaat tcacctggac
     1321 ttcctgaaga gtgaccatcc tgctgtggca cgcatgcgtg tggactcaga caatgcatac
     1381 attggtgtca cctacaaaaa cgaggaagac aagctgaagg actgggaggg tggtctggat
     1441 gagcagagac tgagcgctga cagtggctac atcattcctc tgcctgacat tgaccctgtc
     1501 cctgaggagg aggacctggg caagaggaac agacacagct cgcagacctc tgaagagagt
     1561 gccattgaga cgggttccag cagttccacc ttcatcaaga gagaggacga gaccattgaa
     1621 gacatcgaca tgatggacga catcggcata gactcttcag acctggtgga agacagcttt
     1681 cttaagggtc aagacaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag
     1741 ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     1801 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     1861 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1921 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1981 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2041 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2101 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2161 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2221 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2281 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2341 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2401 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2461 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2521 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2581 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2641 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2701 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2761 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2821 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2881 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     2941 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3001 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3061 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3121 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3181 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3241 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3301 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3361 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3421 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3481 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3541 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3601 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3661 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3721 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     3781 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3841 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3901 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3961 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4021 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4081 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4141 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4201 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4261 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4321 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4381 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4441 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4501 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4561 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4621 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4681 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4741 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4801 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4861 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4921 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4981 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5041 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5101 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5161 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5221 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5281 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5341 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5401 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5461 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5521 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5581 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5641 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5701 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5761 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5821 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5881 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5941 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6001 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6061 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6241 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     6301 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     6361 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     6421 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     6481 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     6541 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6601 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     6661 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     6721 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     6781 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     6841 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     6901 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     6961 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     7021 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
     7081 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     7141 agacccaagc tggctagtta agcttggtac cgagctcgga tccagtaccc ttcaccatgg
     7201 ggacttccca tccggcgttc ctggtcttag gctgtcttct cacagggctg agcctaatcc
     7261 tctgccagct ttcattaccc tctatccttc caaatgaaaa tgaaaaggtt gtgcagctga
     7321 attcatcctt ttctctgaga tgctttgggg agagtgaagt gagctggcag taccccatgt
     7381 ctgaagaaga gagctccgat gtggaaatca gaaatgaaga aaacaacagc ggcctttttg
     7441 tgacggtctt ggaagtgagc agtgcctcgg cggcccacac agggttgtac acttgctatt
     7501 acaaccacac tcagacagaa gagaatgagc ttgaaggcag gcacatttac atctatgtgc
     7561 cagacccaga tgtagccttt gtacctctag gaatgacgga ttatttagtc atcgtggagg
     7621 atgatgattc tgccattata ccttgtcgca caactgatcc cgagactcct gtaaccttac
     7681 acaacagtga gggggtggta cctgcctcct acgacagcag acagggcttt aatgggacct
     7741 tcactgtagg gccctatatc tgtgaggcca ccgtcaaagg aaagaagttc cagaccatcc
     7801 catttaatgt ttatgcttta aaagcaacat cagagctgga tctagaaatg gaagctctta
     7861 aaaccgtgta taagtcaggg gaaacgattg tggtcacctg tgctgttttt aacaatgagg
     7921 tggttgacct tcaatggact taccctggag aagtgaaagg caaaggcatc acaatgctgg
     7981 aagaaatcaa agtcccatcc atcaaattgg tgtacacttt gacggtcccc gaggccacgg
     8041 tgaaagacag tggagattac gaatgtgctg cccgccaggc taccagggag gtcaaagaaa
     8101 tgaagaaagt cactatttct gtccatgaga aaggtttcat tgaaatcaaa cccaccttca
     8161 gccagttgga agctgtcaac ctgcatgaag tcaaacattt tgttgtagag gtgcgggcct
     8221 acccacctcc caggatatcc tggctgaaaa acaatctgac tctgattgaa aatctcactg
     8281 agatcaccac tgatgtggaa aagattcagg aaataaggta tcgaagcaaa ttaaagctga
     8341 tccgtgctaa ggaagaagac agtggccatt atactattgt agctcaaaat gaagatgctg
     8401 tgaagagcta tacttttgaa ctgttaactc aagttccttc atccattctg gacttggtcg
     8461 atgatcacca tggctcaact gggggacaga cggtgaggtg cacagctgaa ggcacgccgc
     8521 ttcctgatat tgagtggatg atatgcaaag atattaagaa atgtaataat gaaacttcct
     8581 ggactatttt ggccaacaat gtctcaaaca tcatcacgga gatccactcc cgagacagga
     8641 gtaccgtgga gggccgtgtg actttcgcca aagtggagga gaccatcgcc gtgcgatgcc
     8701 tggctaagaa tctccttgga gctgagaacc gagagctgaa gctggtggct cccaccctgc
     8761 gttctgaact cacggtggct
//
