LOCUS       sequence-469607-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the D835Y kinase defective mutant variant of human 
            FLT3 that is associated with Acute Myeloid Leukemia and Acute 
            Lymphoblastic 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-D835Y-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3990..4031
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4041..4058
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4081..4098)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4087..4311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4357..4785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4444..4463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4654..4675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4794..4814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4799..5128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4979..5114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5041..5060
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5195..5989
                     /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(5249..5268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5859..5878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6165..6286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6202..6221)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6256..6275
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6335..6351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6335..6351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6348..6370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6359..6375
                     /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(6383..6389,6390..6407,6408..6413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6428..6449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6566..6583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6737..7325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6817..6836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7496..8287,8288..8356))
                     /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     8119..8138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8357..8461)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccagtac ccttcaccat gccagcgttg gcacgcggcg
      961 gcggccagct gccgctgctc gttgtttttt ctgcaatgat atttgggact attacaaatc
     1021 aagatctgcc tgtgatcaag tgtgttttaa tcaatcataa gaacaatgat tcatcagtgg
     1081 ggaagtcatc atcatatccc atggtatcag aatccccgga agacctcggg tgtgcgttga
     1141 gaccccagag ctcagggaca gtgtacgaag ctgccgctgt ggaagtggat gtatctgctt
     1201 ccatcacact gcaagtgctg gtcgatgccc cagggaacat ttcctgtctc tgggtcttta
     1261 agcacagctc cctgaattgc cagccacatt ttgatttaca aaacagagga gttgtttcca
     1321 tggtcatttt gaaaatgaca gaaacccaag ctggagaata cctacttttt attcagagtg
     1381 aagctaccaa ttacacaata ttgtttacag tgagtataag aaataccctg ctttacacat
     1441 taagaagacc ttactttaga aaaatggaaa accaggacgc cctggtctgc atatctgaga
     1501 gcgttccaga gccgatcgtg gaatgggtgc tttgcgattc acagggggaa agctgtaaag
     1561 aagaaagtcc agctgttgtt aaaaaggagg aaaaagtgct tcatgaatta tttgggatgg
     1621 acataaggtg ctgtgccaga aatgaactgg gcagggaatg caccaggctg ttcacaatag
     1681 atctaaatca aactcctcag accacattgc cacaattatt tcttaaagta ggggaaccct
     1741 tatggataag gtgcaaagct gttcatgtga accatggatt cgggctcacc tgggaattag
     1801 aaaacaaagc actcgaggag ggcaactact ttgagatgag tacctattca acaaacagaa
     1861 ctatgatacg gattctgttt gcttttgtat catcagtggc aagaaacgac accggatact
     1921 acacttgttc ctcttcaaag catcccagtc aatcagcttt ggttaccatc gtagaaaagg
     1981 gatttataaa tgctaccaat tcaagtgaag attatgaaat tgaccaatat gaagagtttt
     2041 gtttttctgt caggtttaaa gcctacccac aaatcagatg tacgtggacc ttctctcgaa
     2101 aatcatttcc ttgtgagcaa aagggtcttg ataacggata cagcatatcc aagttttgca
     2161 atcataagca ccagccagga gaatatatat tccatgcaga aaatgatgat gcccaattta
     2221 ccaaaatgtt cacgctgaat ataagaagga aacctcaagt gctcgcagaa gcatcggcaa
     2281 gtcaggcgtc ctgtttctcg gatggatacc cattaccatc ttggacctgg aagaagtgtt
     2341 cagacaagtc tcccaactgc acagaagaga tcacagaagg agtctggaat agaaaggcta
     2401 acagaaaagt gtttggacag tgggtgtcga gcagtactct aaacatgagt gaagccataa
     2461 aagggttcct ggtcaagtgc tgtgcataca attcccttgg cacatcttgt gagacgatcc
     2521 ttttaaactc tccaggcccc ttccctttca tccaagacaa catctcattc tatgcaacaa
     2581 ttggtgtttg tctcctcttc attgtcgttt taaccctgct aatttgtcac aagtacaaaa
     2641 agcaatttag gtatgaaagc cagctacaga tggtacaggt gaccggctcc tcagataatg
     2701 agtacttcta cgttgatttc agagaatatg aatatgatct caaatgggag tttccaagag
     2761 aaaatttaga gtttgggaag gtactaggat caggtgcttt tggaaaagtg atgaacgcaa
     2821 cagcttatgg aattagcaaa acaggagtct caatccaggt tgccgtcaaa atgctgaaag
     2881 aaaaagcaga cagctctgaa agagaggcac tcatgtcaga actcaagatg atgacccagc
     2941 tgggaagcca cgagaatatt gtgaacctgc tgggggcgtg cacactgtca ggaccaattt
     3001 acttgatttt tgaatactgt tgctatggtg atcttctcaa ctatctaaga agtaaaagag
     3061 aaaaatttca caggacttgg acagagattt tcaaggaaca caatttcagt ttttacccca
     3121 ctttccaatc acatccaaat tccagcatgc ctggttcaag agaagttcag atacacccgg
     3181 actcggatca aatctcaggg cttcatggga attcatttca ctctgaagat gaaattgaat
     3241 atgaaaacca aaaaaggctg gaagaagagg aggacttgaa tgtgcttaca tttgaagatc
     3301 ttctttgctt tgcatatcaa gttgccaaag gaatggaatt tctggaattt aagtcgtgtg
     3361 ttcacagaga cctggccgcc aggaacgtgc ttgtcaccca cgggaaagtg gtgaagatat
     3421 gtgactttgg attggctcga tatatcatga gtgattccaa ctatgttgtc aggggcaatg
     3481 cccgtctgcc tgtaaaatgg atggcccccg aaagcctgtt tgaaggcatc tacaccatta
     3541 agagtgatgt ctggtcatat ggaatattac tgtgggaaat cttctcactt ggtgtgaatc
     3601 cttaccctgg cattccggtt gatgctaact tctacaaact gattcaaaat ggatttaaaa
     3661 tggatcagcc attttatgct acagaagaaa tatacattat aatgcaatcc tgctgggctt
     3721 ttgactcaag gaaacggcca tccttcccta atttgacttc gtttttagga tgtcagctgg
     3781 cagatgcaga agaagcgatg tatcagaatg tggatggccg tgtttcggaa tgtcctcaca
     3841 cctaccaaaa caggcgacct ttcagcagag agatggattt ggggctactc tctccgcagg
     3901 ctcaggtcga agattcgaag ggtcaagaca attctgcaga tatccagcac agtggcggcc
     3961 gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     4021 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     4081 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4141 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4201 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4261 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4321 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4381 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4441 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4501 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4561 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4621 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4681 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4741 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4801 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4861 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4921 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4981 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5041 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5101 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5161 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5221 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5281 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5341 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5401 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5461 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5521 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5581 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5641 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5701 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5761 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5821 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5881 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5941 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6001 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6061 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6121 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6301 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6481 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6541 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6601 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7501 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7561 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7621 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7681 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7741 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7801 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7861 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7921 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7981 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8041 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8101 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8161 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8221 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8281 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8341 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8401 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8461 gcacatttcc ccgaaaagtg ccacctgacg tc
//
