LOCUS       sequence-435250-        8447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human KIT receptor tyrosine kinase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8447)
  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 8447)
  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-KIT-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3946..3987
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3997..4014
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4037..4054)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4043..4267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4313..4741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4400..4419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4610..4631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4750..4770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4755..5084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4935..5070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4997..5016
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5151..5945
                     /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(5205..5224)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5815..5834
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6121..6242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6158..6177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6212..6231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6291..6307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6291..6307)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6304..6326)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6315..6331
                     /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(6339..6345,6346..6363,6364..6369))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6384..6405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6522..6539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6693..7281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6773..6792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7452..8243,8244..8312))
                     /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     8075..8094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8313..8417)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 ttaagcttgg taccgagctc ggatccagta cccttcacca tgagaggcgc tcgcggcgcc
      961 tgggattttc tctgcgttct gctcctactg cttcgcgtcc agacaggctc ttctcaacca
     1021 tctgtgagtc caggggaacc gtctccacca tccatccatc caggaaaatc agacttaata
     1081 gtccgcgtgg gcgacgagat taggctgtta tgcactgatc cgggctttgt caaatggact
     1141 tttgagatcc tggatgaaac gaatgagaat aagcagaatg aatggatcac ggaaaaggca
     1201 gaagccacca acaccggcaa atacacgtgc accaacaaac acggcttaag caattccatt
     1261 tatgtgtttg ttagagatcc tgccaagctt ttccttgttg accgctcctt gtatgggaaa
     1321 gaagacaacg acacgctggt ccgctgtcct ctcacagacc cagaagtgac caattattcc
     1381 ctcaaggggt gccaggggaa gcctcttccc aaggacttga ggtttattcc tgaccccaag
     1441 gcgggcatca tgatcaaaag tgtgaaacgc gcctaccatc ggctctgtct gcattgttct
     1501 gtggaccagg agggcaagtc agtgctgtcg gaaaaattca tcctgaaagt gaggccagcc
     1561 ttcaaagctg tgcctgttgt gtctgtgtcc aaagcaagct atcttcttag ggaaggggaa
     1621 gaattcacag tgacgtgcac aataaaagat gtgtctagtt ctgtgtactc aacgtggaaa
     1681 agagaaaaca gtcagcagac taaactacag gagaaatata atagctggca tcacggtgac
     1741 ttcaattatg aacgtcaggc aacgttgact atcagttcag cgagagttaa tgattctgga
     1801 gtgttcatgt gttatgccaa taatactttt ggatcagcaa atgtcacaac aaccttggaa
     1861 gtagtagata aaggattcat taatatcttc cccatgataa acactacagt atttgtaaac
     1921 gatggagaaa atgtagattt gattgttgaa tatgaagcat tccccaaacc tgaacaccag
     1981 cagtggatct atatgaacag aaccttcact gataaatggg aagattatcc caagtctgag
     2041 aatgaaagta atatcagata cgtaagtgaa cttcatctaa cgagattaaa aggcaccgaa
     2101 ggaggcactt acacattcct agtgtccaat tctgacgtca atgctgccat agcatttaat
     2161 gtttatgtga atacaaaacc agaaatcctg acttacgaca ggctcgtgaa tggcatgctc
     2221 caatgtgtgg cagcaggatt cccagagccc acaatagatt ggtatttttg tccaggaact
     2281 gagcagagat gctctgcttc tgtactgcca gtggatgtgc agacactaaa ctcatctggg
     2341 ccaccgtttg gaaagctagt ggttcagagt tctatagatt ctagtgcatt caagcacaat
     2401 ggcacggttg aatgtaaggc ttacaacgat gtgggcaaga cttctgccta ttttaacttt
     2461 gcatttaaag gtaacaacaa agagcaaatc catccccaca ccctgttcac tcctttgctg
     2521 attggtttcg taatcgtagc tggcatgatg tgcattattg tgatgattct gacctacaaa
     2581 tatttacaga aacccatgta tgaagtacag tggaaggttg ttgaggagat aaatggaaac
     2641 aattatgttt acatagaccc aacacaactt ccttatgatc acaaatggga gtttcccaga
     2701 aacaggctga gttttgggaa aaccctgggt gctggagctt tcgggaaggt tgttgaggca
     2761 actgcttatg gcttaattaa gtcagatgcg gccatgactg tcgctgtaaa gatgctcaag
     2821 ccgagtgccc atttgacaga acgggaagcc ctcatgtctg aactcaaagt cctgagttac
     2881 cttggtaatc acatgaatat tgtgaatcta cttggagcct gcaccattgg agggcccacc
     2941 ctggtcatta cagaatattg ttgctatggt gatcttttga attttttgag aagaaaacgt
     3001 gattcattta tttgttcaaa gcaggaagat catgcagaag ctgcacttta taagaatctt
     3061 ctgcattcaa aggagtcttc ctgcagcgat agtactaatg agtacatgga catgaaacct
     3121 ggagtttctt atgttgtccc aaccaaggcc gacaaaagga gatctgtgag aataggctca
     3181 tacatagaaa gagatgtgac tcccgccatc atggaggatg acgagttggc cctagactta
     3241 gaagacttgc tgagcttttc ttaccaggtg gcaaagggca tggctttcct cgcctccaag
     3301 aattgtattc acagagactt ggcagccaga aatatcctcc ttactcatgg tcggatcaca
     3361 aagatttgtg attttggtct agccagagac atcaagaatg attctaatta tgtggttaaa
     3421 ggaaacgctc gactacctgt gaagtggatg gcacctgaaa gcattttcaa ctgtgtatac
     3481 acgtttgaaa gtgacgtctg gtcctatggg atttttcttt gggagctgtt ctctttagga
     3541 agcagcccct atcctggaat gccggtcgat tctaagttct acaagatgat caaggaaggc
     3601 ttccggatgc tcagccctga acacgcacct gctgaaatgt atgacataat gaagacttgc
     3661 tgggatgcag atcccctaaa aagaccaaca ttcaagcaaa ttgttcagct aattgagaag
     3721 cagatttcag agagcaccaa tcatatttac tccaacttag caaactgcag ccccaaccga
     3781 cagaagcccg tggtagacca ttctgtgcgg atcaattctg tcggcagcac cgcttcctcc
     3841 tcccagcctc tgcttgtgca cgacgatgtc gggaagggtc aagacaattc tgcagatatc
     3901 cagcacagtg gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct
     3961 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     4021 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4081 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4141 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4201 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4261 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4321 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4381 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4441 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4501 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4561 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4621 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4681 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4741 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5101 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5161 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5221 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5281 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5341 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5401 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5461 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5521 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5581 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5641 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5701 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5761 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5821 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5881 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5941 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     6001 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6061 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6121 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6181 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6241 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6301 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6361 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6421 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6481 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6541 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6601 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6661 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6721 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6781 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6841 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6901 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6961 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7021 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7081 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7141 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7201 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7261 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7321 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7381 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7441 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7501 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7561 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7621 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7681 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7741 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7801 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7861 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7921 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7981 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8041 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8101 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8161 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8221 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8281 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8341 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8401 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgt
//
