LOCUS       sequence-469587-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the D835N kinase defective mutant variant of human 
            FLT3 that is associated with Acute Myeloid Leukemia.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8492)
  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 8492)
  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-FLT3-D835N-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3989..4030
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4040..4057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4080..4097)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4086..4310
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4356..4784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4443..4462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4653..4674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4793..4813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4798..5127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4978..5113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5040..5059
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5194..5988
                     /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(5248..5267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5858..5877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6164..6285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6201..6220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6255..6274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6334..6350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6334..6350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6347..6369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6358..6374
                     /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(6382..6388,6389..6406,6407..6412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6427..6448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6565..6582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6736..7324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6816..6835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7495..8286,8287..8355))
                     /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     8118..8137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8356..8460)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagtt
      901 aagcttggta ccgagctcgg atccagtacc cttcaccatg ccagcgttgg cacgcggcgg
      961 cggccagctg ccgctgctcg ttgttttttc tgcaatgata tttgggacta ttacaaatca
     1021 agatctgcct gtgatcaagt gtgttttaat caatcataag aacaatgatt catcagtggg
     1081 gaagtcatca tcatatccca tggtatcaga atccccggaa gacctcgggt gtgcgttgag
     1141 accccagagc tcagggacag tgtacgaagc tgccgctgtg gaagtggatg tatctgcttc
     1201 catcacactg caagtgctgg tcgatgcccc agggaacatt tcctgtctct gggtctttaa
     1261 gcacagctcc ctgaattgcc agccacattt tgatttacaa aacagaggag ttgtttccat
     1321 ggtcattttg aaaatgacag aaacccaagc tggagaatac ctacttttta ttcagagtga
     1381 agctaccaat tacacaatat tgtttacagt gagtataaga aataccctgc tttacacatt
     1441 aagaagacct tactttagaa aaatggaaaa ccaggacgcc ctggtctgca tatctgagag
     1501 cgttccagag ccgatcgtgg aatgggtgct ttgcgattca cagggggaaa gctgtaaaga
     1561 agaaagtcca gctgttgtta aaaaggagga aaaagtgctt catgaattat ttgggatgga
     1621 cataaggtgc tgtgccagaa atgaactggg cagggaatgc accaggctgt tcacaataga
     1681 tctaaatcaa actcctcaga ccacattgcc acaattattt cttaaagtag gggaaccctt
     1741 atggataagg tgcaaagctg ttcatgtgaa ccatggattc gggctcacct gggaattaga
     1801 aaacaaagca ctcgaggagg gcaactactt tgagatgagt acctattcaa caaacagaac
     1861 tatgatacgg attctgtttg cttttgtatc atcagtggca agaaacgaca ccggatacta
     1921 cacttgttcc tcttcaaagc atcccagtca atcagctttg gttaccatcg tagaaaaggg
     1981 atttataaat gctaccaatt caagtgaaga ttatgaaatt gaccaatatg aagagttttg
     2041 tttttctgtc aggtttaaag cctacccaca aatcagatgt acgtggacct tctctcgaaa
     2101 atcatttcct tgtgagcaaa agggtcttga taacggatac agcatatcca agttttgcaa
     2161 tcataagcac cagccaggag aatatatatt ccatgcagaa aatgatgatg cccaatttac
     2221 caaaatgttc acgctgaata taagaaggaa acctcaagtg ctcgcagaag catcggcaag
     2281 tcaggcgtcc tgtttctcgg atggataccc attaccatct tggacctgga agaagtgttc
     2341 agacaagtct cccaactgca cagaagagat cacagaagga gtctggaata gaaaggctaa
     2401 cagaaaagtg tttggacagt gggtgtcgag cagtactcta aacatgagtg aagccataaa
     2461 agggttcctg gtcaagtgct gtgcatacaa ttcccttggc acatcttgtg agacgatcct
     2521 tttaaactct ccaggcccct tccctttcat ccaagacaac atctcattct atgcaacaat
     2581 tggtgtttgt ctcctcttca ttgtcgtttt aaccctgcta atttgtcaca agtacaaaaa
     2641 gcaatttagg tatgaaagcc agctacagat ggtacaggtg accggctcct cagataatga
     2701 gtacttctac gttgatttca gagaatatga atatgatctc aaatgggagt ttccaagaga
     2761 aaatttagag tttgggaagg tactaggatc aggtgctttt ggaaaagtga tgaacgcaac
     2821 agcttatgga attagcaaaa caggagtctc aatccaggtt gccgtcaaaa tgctgaaaga
     2881 aaaagcagac agctctgaaa gagaggcact catgtcagaa ctcaagatga tgacccagct
     2941 gggaagccac gagaatattg tgaacctgct gggggcgtgc acactgtcag gaccaattta
     3001 cttgattttt gaatactgtt gctatggtga tcttctcaac tatctaagaa gtaaaagaga
     3061 aaaatttcac aggacttgga cagagatttt caaggaacac aatttcagtt tttaccccac
     3121 tttccaatca catccaaatt ccagcatgcc tggttcaaga gaagttcaga tacacccgga
     3181 ctcggatcaa atctcagggc ttcatgggaa ttcatttcac tctgaagatg aaattgaata
     3241 tgaaaaccaa aaaaggctgg aagaagagga ggacttgaat gtgcttacat ttgaagatct
     3301 tctttgcttt gcatatcaag ttgccaaagg aatggaattt ctggaattta agtcgtgtgt
     3361 tcacagagac ctggccgcca ggaacgtgct tgtcacccac gggaaagtgg tgaagatatg
     3421 tgactttgga ttggctcgaa atatcatgag tgattccaac tatgttgtca ggggcaatgc
     3481 ccgtctgcct gtaaaatgga tggcccccga aagcctgttt gaaggcatct acaccattaa
     3541 gagtgatgtc tggtcatatg gaatattact gtgggaaatc ttctcacttg gtgtgaatcc
     3601 ttaccctggc attccggttg atgctaactt ctacaaactg attcaaaatg gatttaaaat
     3661 ggatcagcca ttttatgcta cagaagaaat atacattata atgcaatcct gctgggcttt
     3721 tgactcaagg aaacggccat ccttccctaa tttgacttcg tttttaggat gtcagctggc
     3781 agatgcagaa gaagcgatgt atcagaatgt ggatggccgt gtttcggaat gtcctcacac
     3841 ctaccaaaac aggcgacctt tcagcagaga gatggatttg gggctactct ctccgcaggc
     3901 tcaggtcgaa gattcgaagg gtcaagacaa ttctgcagat atccagcaca gtggcggccg
     3961 ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct
     4021 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     4081 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4141 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4201 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4261 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4321 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4381 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4441 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4501 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4561 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4621 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4681 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4741 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4801 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4861 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4921 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4981 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5041 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5101 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5161 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5221 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5281 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5341 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5401 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5461 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5521 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5581 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5641 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5701 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5761 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5821 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5881 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5941 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6001 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6061 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6121 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6301 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8461 cacatttccc cgaaaagtgc cacctgacgt cg
//
