LOCUS       sequence-396816-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human PDGFRA receptor tyrosine kinase in mammalian 
            cells.
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-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        229..608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     763..783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     857..876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        857..875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4272..4313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4323..4340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4363..4380)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4369..4593
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4639..5067
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4726..4745)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4936..4957
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5076..5096)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5081..5410
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5261..5396
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5323..5342
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5477..6271
                     /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(5531..5550)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6141..6160
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6447..6568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6484..6503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6538..6557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6617..6633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6617..6633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6630..6652)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6641..6657
                     /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(6665..6671,6672..6689,6690..6695))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6710..6731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6848..6865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7019..7607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7099..7118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7778..8569,8570..8638))
                     /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     8401..8420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8639..8743)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      121 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      181 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      781 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      841 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct
      901 tggtaccgag ctcggatcca gtacccttca ccatggggac ttcccatccg gcgttcctgg
      961 tcttaggctg tcttctcaca gggctgagcc taatcctctg ccagctttca ttaccctcta
     1021 tccttccaaa tgaaaatgaa aaggttgtgc agctgaattc atccttttct ctgagatgct
     1081 ttggggagag tgaagtgagc tggcagtacc ccatgtctga agaagagagc tccgatgtgg
     1141 aaatcagaaa tgaagaaaac aacagcggcc tttttgtgac ggtcttggaa gtgagcagtg
     1201 cctcggcggc ccacacaggg ttgtacactt gctattacaa ccacactcag acagaagaga
     1261 atgagcttga aggcaggcac atttacatct atgtgccaga cccagatgta gcctttgtac
     1321 ctctaggaat gacggattat ttagtcatcg tggaggatga tgattctgcc attatacctt
     1381 gtcgcacaac tgatcccgag actcctgtaa ccttacacaa cagtgagggg gtggtacctg
     1441 cctcctacga cagcagacag ggctttaatg ggaccttcac tgtagggccc tatatctgtg
     1501 aggccaccgt caaaggaaag aagttccaga ccatcccatt taatgtttat gctttaaaag
     1561 caacatcaga gctggatcta gaaatggaag ctcttaaaac cgtgtataag tcaggggaaa
     1621 cgattgtggt cacctgtgct gtttttaaca atgaggtggt tgaccttcaa tggacttacc
     1681 ctggagaagt gaaaggcaaa ggcatcacaa tgctggaaga aatcaaagtc ccatccatca
     1741 aattggtgta cactttgacg gtccccgagg ccacggtgaa agacagtgga gattacgaat
     1801 gtgctgcccg ccaggctacc agggaggtca aagaaatgaa gaaagtcact atttctgtcc
     1861 atgagaaagg tttcattgaa atcaaaccca ccttcagcca gttggaagct gtcaacctgc
     1921 atgaagtcaa acattttgtt gtagaggtgc gggcctaccc acctcccagg atatcctggc
     1981 tgaaaaacaa tctgactctg attgaaaatc tcactgagat caccactgat gtggaaaaga
     2041 ttcaggaaat aaggtatcga agcaaattaa agctgatccg tgctaaggaa gaagacagtg
     2101 gccattatac tattgtagct caaaatgaag atgctgtgaa gagctatact tttgaactgt
     2161 taactcaagt tccttcatcc attctggact tggtcgatga tcaccatggc tcaactgggg
     2221 gacagacggt gaggtgcaca gctgaaggca cgccgcttcc tgatattgag tggatgatat
     2281 gcaaagatat taagaaatgt aataatgaaa cttcctggac tattttggcc aacaatgtct
     2341 caaacatcat cacggagatc cactcccgag acaggagtac cgtggagggc cgtgtgactt
     2401 tcgccaaagt ggaggagacc atcgccgtgc gatgcctggc taagaatctc cttggagctg
     2461 agaaccgaga gctgaagctg gtggctccca ccctgcgttc tgaactcacg gtggctgctg
     2521 cagtcctggt gctgttggtg attgtgatca tctcacttat tgtcctggtt gtcatttgga
     2581 aacagaaacc gaggtatgaa attcgctgga gggtcattga atcaatcagc ccggatggac
     2641 atgaatatat ttatgtggac ccgatgcagc tgccttatga ctcaagatgg gagtttccaa
     2701 gagatggact agtgcttggt cgggtcttgg ggtctggagc gtttgggaag gtggttgaag
     2761 gaacagccta tggattaagc cggtcccaac ctgtcatgaa agttgcagtg aagatgctaa
     2821 aacccacggc cagatccagt gaaaaacaag ctctcatgtc tgaactgaag ataatgactc
     2881 acctggggcc acatttgaac attgtaaact tgctgggagc ctgcaccaag tcaggcccca
     2941 tttacatcat cacagagtat tgcttctatg gagatttggt caactatttg cataagaata
     3001 gggatagctt cctgagccac cacccagaga agccaaagaa agagctggat atctttggat
     3061 tgaaccctgc tgatgaaagc acacggagct atgttatttt atcttttgaa aacaatggtg
     3121 actacatgga catgaagcag gctgatacta cacagtatgt ccccatgcta gaaaggaaag
     3181 aggtttctaa atattccgac atccagagat cactctatga tcgtccagcc tcatataaga
     3241 agaaatctat gttagactca gaagtcaaaa acctcctttc agatgataac tcagaaggcc
     3301 ttactttatt ggatttgttg agcttcacct atcaagttgc ccgaggaatg gagtttttgg
     3361 cttcaaaaaa ttgtgtccac cgtgatctgg ctgctcgcaa cgtcctcctg gcacaaggaa
     3421 aaattgtgaa gatctgtgac tttggcctgg ccagagacat catgcatgat tcgaactatg
     3481 tgtcgaaagg cagtaccttt ctgcccgtga agtggatggc tcctgagagc atctttgaca
     3541 acctctacac cacactgagt gatgtctggt cttatggcat tctgctctgg gagatctttt
     3601 cccttggtgg caccccttac cccggcatga tggtggattc tactttctac aataagatca
     3661 agagtgggta ccggatggcc aagcctgacc acgctaccag tgaagtctac gagatcatgg
     3721 tgaaatgctg gaacagtgag ccggagaaga gaccctcctt ttaccacctg agtgagattg
     3781 tggagaatct gctgcctgga caatataaaa agagttatga aaaaattcac ctggacttcc
     3841 tgaagagtga ccatcctgct gtggcacgca tgcgtgtgga ctcagacaat gcatacattg
     3901 gtgtcaccta caaaaacgag gaagacaagc tgaaggactg ggagggtggt ctggatgagc
     3961 agagactgag cgctgacagt ggctacatca ttcctctgcc tgacattgac cctgtccctg
     4021 aggaggagga cctgggcaag aggaacagac acagctcgca gacctctgaa gagagtgcca
     4081 ttgagacggg ttccagcagt tccaccttca tcaagagaga ggacgagacc attgaagaca
     4141 tcgacatgat ggacgacatc ggcatagact cttcagacct ggtggaagac agctttctta
     4201 agggtcaaga caattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc
     4261 cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     4321 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     4381 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4441 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4501 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4561 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4621 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4681 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4741 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4801 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4861 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4921 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4981 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5041 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     5101 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5161 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5221 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5281 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5341 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5401 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5461 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5521 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5581 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5641 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5701 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5761 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5821 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5881 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5941 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6001 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6061 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6121 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6181 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6241 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     6301 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6361 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6421 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6541 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6601 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6661 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6721 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6781 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6841 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6901 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6961 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7021 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7081 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7141 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7201 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7261 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7321 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7381 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7441 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7501 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7561 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7621 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7681 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7741 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7801 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7861 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7921 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7981 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8041 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8101 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8161 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8221 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8281 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8341 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8401 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8461 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8521 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8581 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8641 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8701 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8761 tgccacctga cgtcgacgga
//
