LOCUS       sequence-468863-        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"
     CDS             1836..1877
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1887..1904
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1927..1944)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1933..2157
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2203..2631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2290..2309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2500..2521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2640..2660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2645..2974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2825..2960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2887..2906
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3041..3835
                     /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(3095..3114)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3705..3724
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4011..4132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4048..4067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4102..4121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4181..4197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4181..4197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4194..4216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4205..4221
                     /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(4229..4235,4236..4253,4254..4259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4274..4295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4412..4429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4583..5171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4663..4682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5342..6133,6134..6202))
                     /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     5965..5984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6203..6307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6382..6401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6573..6952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6953..7156
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7107..7127
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7201..7220
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7201..7219
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tggctaagaa tctccttgga gctgagaacc gagagctgaa gctggtggct cccaccctgc
       61 gttctgaact cacggtggct gctgcagtcc tggtgctgtt ggtgattgtg atcatctcac
      121 ttattgtcct ggttgtcatt tggaaacaga aaccgaggta tgaaattcgc tggagggtca
      181 ttgaatcaat cagcccggat ggacatgaat atatttatgt ggacccgatg cagctgcctt
      241 atgactcaag atgggagttt ccaagagatg gactagtgct tggtcgggtc ttggggtctg
      301 gagcgtttgg gaaggtggtt gaaggaacag cctatggatt aagccggtcc caacctgtca
      361 tgaaagttgc agtgaagatg ctaaaaccca cggccagatc cagtgaaaaa caagctctca
      421 tgtctgaact gaagataatg actcacctgg ggccacattt gaacattgta aacttgctgg
      481 gagcctgcac caagtcaggc cccatttaca tcatcacaga gtattgcttc tatggagatt
      541 tggtcaacta tttgcataag aatagggata gcttcctgag ccaccaccca gagaagccaa
      601 agaaagagct ggatatcttt ggattgaacc ctgctgatga aagcacacgg agctatgtta
      661 ttttatcttt tgaaaacaat ggtgactaca tggacatgaa gcaggctgat actacacagt
      721 atgtccccat gctagaaagg aaagaggttt ctaaatattc cgacatccag agatcactct
      781 atgatcgtcc agcctcatat aagaagaaat ctatgttaga ctcagaagtc aaaaacctcc
      841 tttcagatga taactcagaa ggccttactt tattggattt gttgagcttc acctatcaag
      901 ttgcccgagg aatggagttt ttggcttcaa aaaattgtgt ccaccgtgat ctggctgctc
      961 gcaacgtcct cctggcacaa ggaaaaattg tgaagatctg tgactttggc ctggccagag
     1021 acatcatgca tgattcgaac tatgtgtcga aaggcagtac ctttctgccc gtgaagtgga
     1081 tggctcctga gagcatcttt gacaacctct acaccacact gagtgatgtc tggtcttatg
     1141 gcattctgct ctgggagatc ttttcccttg gtggcacccc ttaccccggc atgatggtgg
     1201 attctacttt ctacaataag atcaagagtg ggtaccggat ggccaagcct gaccacgcta
     1261 ccagtgaagt ctacgagatc atggtgaaat gctggaacag tgagccggag aagagaccct
     1321 ccttttacca cctgagtgag attgtggaga atctgctgcc tggacaatat aaaaagagtt
     1381 atgaaaaaat tcacctggac ttcctgaaga gtgaccatcc tgctgtggca cgcatgcgtg
     1441 tggactcaga caatgcatac attggtgtca cctacaaaaa cgaggaagac aagctgaagg
     1501 actgggaggg tggtctggat gagcagagac tgagcgctga cagtggctac atcattcctc
     1561 tgcctgacat tgaccctgtc cctgaggagg aggacctggg caagaggaac agacacagct
     1621 cgcagacctc tgaagagagt gccattgaga cgggttccag cagttccacc ttcatcaaga
     1681 gagaggacga gaccattgaa gacatcgaca tgatggacga catcggcata gactcttcag
     1741 acctggtgga agacagcttt cttaagggtc aagacaattc tgcagatatc cagcacagtg
     1801 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     1861 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     1921 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1981 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2041 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2101 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2161 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2401 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2521 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2641 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2701 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2821 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2881 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2941 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3001 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3061 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3121 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3181 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3241 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3301 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3361 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3421 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3481 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3541 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3601 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3661 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3721 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3781 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3841 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4021 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4081 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4141 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4201 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4261 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4321 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4381 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4441 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4501 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4981 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6301 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6361 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6421 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6481 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6541 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     6601 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6661 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6721 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6781 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6841 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6901 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     6961 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     7021 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     7081 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     7141 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     7201 taatacgact cactataggg agacccaagc tggctagtta agcttggtac cgagctcgga
     7261 tccagtaccc ttcaccatgg ggacttccca tccggcgttc ctggtcttag gctgtcttct
     7321 cacagggctg agcctaatcc tctgccagct ttcattaccc tctatccttc caaatgaaaa
     7381 tgaaaaggtt gtgcagctga attcatcctt ttctctgaga tgctttgggg agagtgaagt
     7441 gagctggcag taccccatgt ctgaagaaga gagctccgat gtggaaatca gaaatgaaga
     7501 aaacaacagc ggcctttttg tgacggtctt ggaagtgagc agtgcctcgg cggcccacac
     7561 agggttgtac acttgctatt acaaccacac tcagacagaa gagaatgagc ttgaaggcag
     7621 gcacatttac atctatgtgc cagacccaga tgtagccttt gtacctctag gaatgacgga
     7681 ttatttagtc atcgtggagg atgatgattc tgccattata ccttgtcgca caactgatcc
     7741 cgagactcct gtaaccttac acaacagtga gggggtggta cctgcctcct acgacagcag
     7801 acagggcttt aatgggacct tcactgtagg gccctatatc tgtgaggcca ccgtcaaagg
     7861 aaagaagttc cagaccatcc catttaatgt ttatgcttta aaagcaacat cagagctgga
     7921 tctagaaatg gaagctctta aaaccgtgta taagtcaggg gaaacgattg tggtcacctg
     7981 tgctgttttt aacaatgagg tggttgacct tcaatggact taccctggag aagtgaaagg
     8041 caaaggcatc acaatgctgg aagaaatcaa agtcccatcc atcaaattgg tgtacacttt
     8101 gacggtcccc gaggccacgg tgaaagacag tggagattac gaatgtgctg cccgccaggc
     8161 taccagggag gtcaaagaaa tgaagaaagt cactatttct gtccatgaga aaggtttcat
     8221 tgaaatcaaa cccaccttca gccagttgga agctgtcaac ctgcatgaag tcaaacattt
     8281 tgttgtagag gtgcgggcct acccacctcc caggatatcc tggctgaaaa acaatctgac
     8341 tctgattgaa aatctcactg agatcaccac tgatgtggaa aagattcagg aaataaggta
     8401 tcgaagcaaa ttaaagctga tccgtgctaa ggaagaagac agtggccatt atactattgt
     8461 agctcaaaat gaagatgctg tgaagagcta tacttttgaa ctgttaactc aagttccttc
     8521 atccattctg gacttggtcg atgatcacca tggctcaact gggggacaga cggtgaggtg
     8581 cacagctgaa ggcacgccgc ttcctgatat tgagtggatg atatgcaaag atattaagaa
     8641 atgtaataat gaaacttcct ggactatttt ggccaacaat gtctcaaaca tcatcacgga
     8701 gatccactcc cgagacagga gtaccgtgga gggccgtgtg actttcgcca aagtggagga
     8761 gaccatcgcc gtgcgatgcc
//
