LOCUS       sequence-468954-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the P577S 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-P577S-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     494..513
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        588..692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(693..761,762..1553)
                     /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     complement(911..930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1724..2312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2213..2232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2466..2483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2600..2621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2636..2641,2642..2659,2660..2666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2674..2690
                     /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)."
     primer_bind     2679..2701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2698..2714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2698..2714
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2763..2884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2828..2847
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3060..3854)
                     /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(3171..3190)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3781..3800
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3921..4250)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3935..4070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3989..4008)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4235..4255
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4264..4692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4374..4395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4586..4605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4738..4962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4951..4968
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4991..5008)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5018..5059)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(8455..8474)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8456..8474)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8519..8722)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8548..8568)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(8723..8780,1..322)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
       61 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
      121 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
      181 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
      241 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
      301 taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca
      361 aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta
      421 gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac
      481 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag gggatcggga
      541 gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
      601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
      661 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
      721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
      781 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
      841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
      901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
      961 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1081 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1261 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1321 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1561 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1621 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     1681 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1801 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1861 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1921 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1981 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2041 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2101 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2161 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2221 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2281 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2341 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2401 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2461 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     2521 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     2581 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     2641 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     2701 gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat
     2761 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2821 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2881 agttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     2941 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     3001 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gcgaacccca gagtcccgct
     3061 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     3121 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     3181 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     3241 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     3301 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     3361 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     3421 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     3481 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     3541 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     3601 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     3661 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     3721 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     3781 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     3841 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc gaaaatggat
     3901 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     3961 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
     4021 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     4081 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     4141 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     4201 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
     4261 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4321 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4381 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4441 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4501 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4561 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4621 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4681 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca
     4741 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     4801 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     4861 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     4921 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg
     4981 tttaaactca atggtgatgg tgatgatgac cggtacgcgt agaatcgaga ccgaggagag
     5041 ggttagggat aggcttacct tcgaaccgcg ggccctctag actcgagcgg ccgccactgt
     5101 gctggatatc tgcagaattg tcttgaccct taagaaagct gtcttccacc aggtctgaag
     5161 agtctatgcc gatgtcgtcc atcatgtcga tgtcttcaat ggtctcgtcc tctctcttga
     5221 tgaaggtgga actgctggaa cccgtctcaa tggcactctc ttcagaggtc tgcgagctgt
     5281 gtctgttcct cttgcccagg tcctcctcct cagggacagg gtcaatgtca ggcagaggaa
     5341 tgatgtagcc actgtcagcg ctcagtctct gctcatccag accaccctcc cagtccttca
     5401 gcttgtcttc ctcgtttttg taggtgacac caatgtatgc attgtctgag tccacacgca
     5461 tgcgtgccac agcaggatgg tcactcttca ggaagtccag gtgaattttt tcataactct
     5521 ttttatattg tccaggcagc agattctcca caatctcact caggtggtaa aaggagggtc
     5581 tcttctccgg ctcactgttc cagcatttca ccatgatctc gtagacttca ctggtagcgt
     5641 ggtcaggctt ggccatccgg tacccactct tgatcttatt gtagaaagta gaatccacca
     5701 tcatgccggg gtaaggggtg ccaccaaggg aaaagatctc ccagagcaga atgccataag
     5761 accagacatc actcagtgtg gtgtagaggt tgtcaaagat gctctcagga gccatccact
     5821 tcacgggcag aaaggtactg cctttcgaca catagttcga atcatgcatg atgtctctgg
     5881 ccaggccaaa gtcacagatc ttcacaattt ttccttgtgc caggaggacg ttgcgagcag
     5941 ccagatcacg gtggacacaa ttttttgaag ccaaaaactc cattcctcgg gcaacttgat
     6001 aggtgaagct caacaaatcc aataaagtaa ggccttctga gttatcatct gaaaggaggt
     6061 ttttgacttc tgagtctaac atagatttct tcttatatga ggctggacga tcatagagtg
     6121 atctctggat gtcggaatat ttagaaacct ctttcctttc tagcatgggg acatactgtg
     6181 tagtatcagc ctgcttcatg tccatgtagt caccattgtt ttcaaaagat aaaataacat
     6241 agctccgtgt gctttcatca gcagggttca atccaaagat atccagctct ttctttggct
     6301 tctctgggtg gtggctcagg aagctatccc tattcttatg caaatagttg accaaatctc
     6361 catagaagca atactctgtg atgatgtaaa tggggcctga cttggtgcag gctcccagca
     6421 agtttacaat gttcaaatgt ggccccaggt gagtcattat cttcagttca gacatgagag
     6481 cttgtttttc actggatctg gccgtgggtt ttagcatctt cactgcaact ttcatgacag
     6541 gttgggaccg gcttaatcca taggctgttc cttcaaccac cttcccaaac gctccagacc
     6601 ccaagacccg accaagcact agtccatctc ttggaaactc ccatcttgag tcataaggca
     6661 gctgcatcga gtccacataa atatattcat gtccatccgg gctgattgat tcaatgaccc
     6721 tccagcgaat ttcatacctc ggtttctgtt tccaaatgac aaccaggaca ataagtgaga
     6781 tgatcacaat caccaacagc accaggactg cagcagccac cgtgagttca gaacgcaggg
     6841 tgggagccac cagcttcagc tctcggttct cagctccaag gagattctta gccaggcatc
     6901 gcacggcgat ggtctcctcc actttggcga aagtcacacg gccctccacg gtactcctgt
     6961 ctcgggagtg gatctccgtg atgatgtttg agacattgtt ggccaaaata gtccaggaag
     7021 tttcattatt acatttctta atatctttgc atatcatcca ctcaatatca ggaagcggcg
     7081 tgccttcagc tgtgcacctc accgtctgtc ccccagttga gccatggtga tcatcgacca
     7141 agtccagaat ggatgaagga acttgagtta acagttcaaa agtatagctc ttcacagcat
     7201 cttcattttg agctacaata gtataatggc cactgtcttc ttccttagca cggatcagct
     7261 ttaatttgct tcgatacctt atttcctgaa tcttttccac atcagtggtg atctcagtga
     7321 gattttcaat cagagtcaga ttgtttttca gccaggatat cctgggaggt gggtaggccc
     7381 gcacctctac aacaaaatgt ttgacttcat gcaggttgac agcttccaac tggctgaagg
     7441 tgggtttgat ttcaatgaaa cctttctcat ggacagaaat agtgactttc ttcatttctt
     7501 tgacctccct ggtagcctgg cgggcagcac attcgtaatc tccactgtct ttcaccgtgg
     7561 cctcggggac cgtcaaagtg tacaccaatt tgatggatgg gactttgatt tcttccagca
     7621 ttgtgatgcc tttgcctttc acttctccag ggtaagtcca ttgaaggtca accacctcat
     7681 tgttaaaaac agcacaggtg accacaatcg tttcccctga cttatacacg gttttaagag
     7741 cttccatttc tagatccagc tctgatgttg cttttaaagc ataaacatta aatgggatgg
     7801 tctggaactt ctttcctttg acggtggcct cacagatata gggccctaca gtgaaggtcc
     7861 cattaaagcc ctgtctgctg tcgtaggagg caggtaccac cccctcactg ttgtgtaagg
     7921 ttacaggagt ctcgggatca gttgtgcgac aaggtataat ggcagaatca tcatcctcca
     7981 cgatgactaa ataatccgtc attcctagag gtacaaaggc tacatctggg tctggcacat
     8041 agatgtaaat gtgcctgcct tcaagctcat tctcttctgt ctgagtgtgg ttgtaatagc
     8101 aagtgtacaa ccctgtgtgg gccgccgagg cactgctcac ttccaagacc gtcacaaaaa
     8161 ggccgctgtt gttttcttca tttctgattt ccacatcgga gctctcttct tcagacatgg
     8221 ggtactgcca gctcacttca ctctccccaa agcatctcag agaaaaggat gaattcagct
     8281 gcacaacctt ttcattttca tttggaagga tagagggtaa tgaaagctgg cagaggatta
     8341 ggctcagccc tgtgagaaga cagcctaaga ccaggaacgc cggatgggaa gtccccatgg
     8401 tgaagggtac tggatccgag ctcggtacca agcttaacta gccagcttgg gtctccctat
     8461 agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag
     8521 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     8581 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     8641 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     8701 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     8761 aagtaggaaa gtcccataag
//
