LOCUS       sequence-396815-        8831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human PDGFRB receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8831)
  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 8831)
  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-PDGFRB-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        221..600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        601..804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     755..775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     849..868
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        849..867
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2461..2607
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             4315..4356
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4366..4383
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4406..4423)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4412..4636
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4682..5110
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4769..4788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4979..5000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5119..5139)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5124..5453
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5304..5439
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5366..5385
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5520..6314
                     /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(5574..5593)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6184..6203
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6490..6611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6527..6546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6581..6600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6660..6676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6660..6676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6673..6695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6684..6700
                     /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(6708..6714,6715..6732,6733..6738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6753..6774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6891..6908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7062..7650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7821..8612,8613..8681))
                     /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     8444..8463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8682..8786)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
       61 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      121 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      181 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      241 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      301 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      361 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      421 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      481 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      541 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      601 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      661 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      721 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      781 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      841 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag cttggtaccg
      901 agctcggatc cagtaccctt caccatgcgg cttccgggtg cgatgccagc tctggccctc
      961 aaaggcgagc tgctgttgct gtctctcctg ttacttctgg aaccacagat ctctcagggc
     1021 ctggtcgtca cacccccggg gccagagctt gtcctcaatg tctccagcac cttcgttctg
     1081 acctgctcgg gttcagctcc ggtggtgtgg gaacggatgt cccaggagcc cccacaggaa
     1141 atggccaagg cccaggatgg caccttctcc agcgtgctca cactgaccaa cctcactggg
     1201 ctagacacgg gagaatactt ttgcacccac aatgactccc gtggactgga gaccgatgag
     1261 cggaaacggc tctacatctt tgtgccagat cccaccgtgg gcttcctccc taatgatgcc
     1321 gaggaactat tcatctttct cacggaaata actgagatca ccattccatg ccgagtaaca
     1381 gacccacagc tggtggtgac actgcacgag aagaaagggg acgttgcact gcctgtcccc
     1441 tatgatcacc aacgtggctt tagtggtatc tttgaggaca gaagctacat ctgcaaaacc
     1501 accattgggg acagggaggt ggattctgat gcctactatg tctacagact ccaggtgtca
     1561 tccatcaacg tctctgtgaa cgcagtgcag actgtggtcc gccagggtga gaacatcacc
     1621 ctcatgtgca ttgtgatcgg gaatgaggtg gtcaacttcg agtggacata cccccgcaaa
     1681 gaaagtgggc ggctggtgga gccggtgact gacttcctct tggatatgcc ttaccacatc
     1741 cgctccatcc tgcacatccc cagtgccgag ttagaagact cggggaccta cacctgcaat
     1801 gtgacggaga gtgtgaatga ccatcaggat gaaaaggcca tcaacatcac cgtggttgag
     1861 agcggctacg tgcggctcct gggagaggtg ggcacactac aatttgctga gctgcatcgg
     1921 agccggacac tgcaggtagt gttcgaggcc tacccaccgc ccactgtcct gtggttcaaa
     1981 gacaaccgca ccctgggcga ctccagcgct ggcgaaatcg ccctgtccac gcgcaacgtg
     2041 tcggagaccc ggtatgtgtc agagctgaca ctggttcgcg tgaaggtggc agaggctggc
     2101 cactacacca tgcgggcctt ccatgaggat gctgaggtcc agctctcctt ccagctacag
     2161 atcaatgtcc ctgtccgagt gctggagcta agtgagagcc accctgacag tggggaacag
     2221 acagtccgct gtcgtggccg gggcatgccc cagccgaaca tcatctggtc tgcctgcaga
     2281 gacctcaaaa ggtgtccacg tgagctgccg cccacgctgc tggggaacag ttccgaagag
     2341 gagagccagc tggagactaa cgtgacgtac tgggaggagg agcaggagtt tgaggtggtg
     2401 agcacactgc gtctgcagca cgtggatcgg ccactgtcgg tgcgctgcac gctgcgcaac
     2461 gctgtgggcc aggacacgca ggaggtcatc gtggtgccac actccttgcc ctttaaggtg
     2521 gtggtgatct cagccatcct ggccctggtg gtgctcacca tcatctccct tatcatcctc
     2581 atcatgcttt ggcagaagaa gccacgttac gagatccgat ggaaggtgat tgagtctgtg
     2641 agctctgacg gccatgagta catctacgtg gaccccatgc agctgcccta tgactccacg
     2701 tgggagctgc cgcgggacca gcttgtgctg ggacgcaccc tcggctctgg ggcctttggg
     2761 caggtggtgg aggccacggc tcatggcctg agccattctc aggccacgat gaaagtggcc
     2821 gtcaagatgc ttaaatccac agcccgcagc agtgagaagc aagcccttat gtcggagctg
     2881 aagatcatga gtcaccttgg gccccacctg aacgtggtca acctgttggg ggcctgcacc
     2941 aaaggaggac ccatctatat catcactgag tactgccgct acggagacct ggtggactac
     3001 ctgcaccgca acaaacacac cttcctgcag caccactccg acaagcgccg cccgcccagc
     3061 gcggagctct acagcaatgc tctgcccgtt gggctccccc tgcccagcca tgtgtccttg
     3121 accggggaga gcgacggtgg ctacatggac atgagcaagg acgagtcggt ggactatgtg
     3181 cccatgctgg acatgaaagg agacgtcaaa tatgcagaca tcgagtcctc caactacatg
     3241 gccccttacg ataactacgt tccctctgcc cctgagagga cctgccgagc aactttgatc
     3301 aacgagtctc cagtgctaag ctacatggac ctcgtgggct tcagctacca ggtggccaat
     3361 ggcatggagt ttctggcctc caagaactgc gtccacagag acctggcggc taggaacgtg
     3421 ctcatctgtg aaggcaagct ggtcaagatc tgtgactttg gcctggctcg agacatcatg
     3481 cgggactcga attacatctc caaaggcagc acctttttgc ctttaaagtg gatggctccg
     3541 gagagcatct tcaacagcct ctacaccacc ctgagcgacg tgtggtcctt cgggatcctg
     3601 ctctgggaga tcttcacctt gggtggcacc ccttacccag agctgcccat gaacgagcag
     3661 ttctacaatg ccatcaaacg gggttaccgc atggcccagc ctgcccatgc ctccgacgag
     3721 atctatgaga tcatgcagaa gtgctgggaa gagaagtttg agattcggcc ccccttctcc
     3781 cagctggtgc tgcttctcga gagactgttg ggcgaaggtt acaaaaagaa gtaccagcag
     3841 gtggatgagg agtttctgag gagtgaccac ccagccatcc ttcggtccca ggcccgcttg
     3901 cctgggttcc atggcctccg atctcccctg gacaccagct ccgtcctcta tactgccgtg
     3961 cagcccaatg agggtgacaa cgactatatc atccccctgc ctgaccccaa acccgaggtt
     4021 gctgacgagg gcccactgga gggttccccc agcctagcca gctccaccct gaatgaagtc
     4081 aacacctcct caaccatctc ctgtgacagc cccctggagc cccaggacga accagagcca
     4141 gagccccagc ttgagctcca ggtggagccg gagccagagc tggaacagtt gccggattcg
     4201 gggtgccctg cgcctcgggc ggaagcagag gatagctttc taaagggtca agacaattct
     4261 gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgcggtt cgaaggtaag
     4321 cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac
     4381 cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4441 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4501 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4561 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4621 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4681 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4741 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4801 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4861 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4921 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4981 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5041 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5101 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5221 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5281 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5341 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     5401 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5461 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5521 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5581 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5641 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5701 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5761 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5821 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5881 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5941 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6001 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     6061 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6121 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6181 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6241 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6301 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     6361 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6421 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6481 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6601 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7141 taatgatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8821 ggatcgggag a
//
