LOCUS       sequence-396813-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human FLT3 receptor tyrosine kinase in mammalian 
            cells.
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-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..47)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     847..866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        847..865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3974..4015
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4025..4042
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4065..4082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4071..4295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4341..4769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4638..4659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4778..4798)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4783..5112
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4963..5098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5025..5044
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5179..5973
                     /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(5233..5252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5843..5862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6149..6270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6186..6205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6240..6259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6319..6335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6319..6335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6332..6354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6343..6359
                     /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(6367..6373,6374..6391,6392..6397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6412..6433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6550..6567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6721..7309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6801..6820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7480..8271,8272..8340))
                     /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     8103..8122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8341..8445)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
       61 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      121 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      181 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      841 cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct tggtaccgag
      901 ctcggatcca gtacccttca ccatgccagc gttggcacgc ggcggcggcc agctgccgct
      961 gctcgttgtt ttttctgcaa tgatatttgg gactattaca aatcaagatc tgcctgtgat
     1021 caagtgtgtt ttaatcaatc ataagaacaa tgattcatca gtggggaagt catcatcata
     1081 tcccatggta tcagaatccc cggaagacct cgggtgtgcg ttgagacccc agagctcagg
     1141 gacagtgtac gaagctgccg ctgtggaagt ggatgtatct gcttccatca cactgcaagt
     1201 gctggtcgat gccccaggga acatttcctg tctctgggtc tttaagcaca gctccctgaa
     1261 ttgccagcca cattttgatt tacaaaacag aggagttgtt tccatggtca ttttgaaaat
     1321 gacagaaacc caagctggag aatacctact ttttattcag agtgaagcta ccaattacac
     1381 aatattgttt acagtgagta taagaaatac cctgctttac acattaagaa gaccttactt
     1441 tagaaaaatg gaaaaccagg acgccctggt ctgcatatct gagagcgttc cagagccgat
     1501 cgtggaatgg gtgctttgcg attcacaggg ggaaagctgt aaagaagaaa gtccagctgt
     1561 tgttaaaaag gaggaaaaag tgcttcatga attatttggg acggacataa ggtgctgtgc
     1621 cagaaatgaa ctgggcaggg aatgcaccag gctgttcaca atagatctaa atcaaactcc
     1681 tcagaccaca ttgccacaat tatttcttaa agtaggggaa cccttatgga taaggtgcaa
     1741 agctgttcat gtgaaccatg gattcgggct cacctgggaa ttagaaaaca aagcactcga
     1801 ggagggcaac tactttgaga tgagtaccta ttcaacaaac agaactatga tacggattct
     1861 gtttgctttt gtatcatcag tggcaagaaa cgacaccgga tactacactt gttcctcttc
     1921 aaagcatccc agtcaatcag ctttggttac catcgtagaa aagggattta taaatgctac
     1981 caattcaagt gaagattatg aaattgacca atatgaagag ttttgttttt ctgtcaggtt
     2041 taaagcctac ccacaaatca gatgtacgtg gaccttctct cgaaaatcat ttccttgtga
     2101 gcaaaagggt cttgataacg gatacagcat atccaagttt tgcaatcata agcaccagcc
     2161 aggagaatat atattccatg cagaaaatga tgatgcccaa tttaccaaaa tgttcacgct
     2221 gaatataaga aggaaacctc aagtgctcgc agaagcatcg gcaagtcagg cgtcctgttt
     2281 ctcggatgga tacccattac catcttggac ctggaagaag tgttcagaca agtctcccaa
     2341 ctgcacagaa gagatcacag aaggagtctg gaatagaaag gctaacagaa aagtgtttgg
     2401 acagtgggtg tcgagcagta ctctaaacat gagtgaagcc ataaaagggt tcctggtcaa
     2461 gtgctgtgca tacaattccc ttggcacatc ttgtgagacg atccttttaa actctccagg
     2521 ccccttccct ttcatccaag acaacatctc attctatgca acaattggtg tttgtctcct
     2581 cttcattgtc gttttaaccc tgctaatttg tcacaagtac aaaaagcaat ttaggtatga
     2641 aagccagcta cagatggtac aggtgaccgg ctcctcagat aatgagtact tctacgttga
     2701 tttcagagaa tatgaatatg atctcaaatg ggagtttcca agagaaaatt tagagtttgg
     2761 gaaggtacta ggatcaggtg cttttggaaa agtgatgaac gcaacagctt atggaattag
     2821 caaaacagga gtctcaatcc aggttgccgt caaaatgctg aaagaaaaag cagacagctc
     2881 tgaaagagag gcactcatgt cagaactcaa gatgatgacc cagctgggaa gccacgagaa
     2941 tattgtgaac ctgctggggg cgtgcacact gtcaggacca atttacttga tttttgaata
     3001 ctgttgctat ggtgatcttc tcaactatct aagaagtaaa agagaaaaat ttcacaggac
     3061 ttggacagag attttcaagg aacacaattt cagtttttac cccactttcc aatcacatcc
     3121 aaattccagc atgcctggtt caagagaagt tcagatacac ccggactcgg atcaaatctc
     3181 agggcttcat gggaattcat ttcactctga agatgaaatt gaatatgaaa accaaaaaag
     3241 gctggaagaa gaggaggact tgaatgtgct tacatttgaa gatcttcttt gctttgcata
     3301 tcaagttgcc aaaggaatgg aatttctgga atttaagtcg tgtgttcaca gagacctggc
     3361 cgccaggaac gtgcttgtca cccacgggaa agtggtgaag atatgtgact ttggattggc
     3421 tcgagatatc atgagtgatt ccaactatgt tgtcaggggc aatgcccgtc tgcctgtaaa
     3481 atggatggcc cccgaaagcc tgtttgaagg catctacacc attaagagtg atgtctggtc
     3541 atatggaata ttactgtggg aaatcttctc acttggtgtg aatccttacc ctggcattcc
     3601 ggttgatgct aacttctaca aactgattca aaatggattt aaaatggatc agccatttta
     3661 tgctacagaa gaaatataca ttataatgca atcctgctgg gcttttgact caaggaaacg
     3721 gccatccttc cctaatttga cttcgttttt aggatgtcag ctggcagatg cagaagaagc
     3781 gatgtatcag aatgtggatg gccgtgtttc ggaatgtcct cacacctacc aaaacaggcg
     3841 acctttcagc agagagatgg atttggggct actctctccg caggctcagg tcgaagattc
     3901 gaagggtcaa gacaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg
     3961 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     4021 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     4081 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4141 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4201 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4261 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4321 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4741 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4801 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4861 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4921 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4981 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5041 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5101 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5161 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5221 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5281 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5341 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5401 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5461 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5521 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5581 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5641 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5701 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5761 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5821 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5881 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5941 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6001 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6061 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6121 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6241 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6301 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6361 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6421 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6481 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6541 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6601 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6661 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6721 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6781 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6841 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6901 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6961 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7021 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7081 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7141 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7201 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7261 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7321 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7381 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7441 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7501 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7561 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7621 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7681 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7741 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7801 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7861 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7921 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7981 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8041 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8101 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8161 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8221 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8281 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8341 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8401 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8461 agtgccacct gacgtcgacg gatcgggaga tc
//
