LOCUS       sequence-407728-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TRKB receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  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 7979)
  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-TRKB-V5/HIS
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(129..233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(308..327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        499..878
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        879..1082
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1033..1053
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1127..1146
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1127..1145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3741..3782
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3792..3809
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3832..3849)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3838..4062
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4108..4536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4195..4214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4405..4426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4545..4565)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4550..4879
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4730..4865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4792..4811
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4946..5740
                     /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(5000..5019)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5610..5629
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5916..6037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5953..5972)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6007..6026
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6086..6102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6086..6102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6099..6121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6110..6126
                     /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(6134..6140,6141..6158,6159..6164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6179..6200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6317..6334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6488..7076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6568..6587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7247..7979,1..59,60..128))
                     /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     7870..7889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
       61 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      121 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      181 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      241 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
      301 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      361 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      421 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      481 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      541 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      601 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      661 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      721 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      781 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      841 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      901 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      961 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1021 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1081 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     1141 atagggagac ccaagctggc tagttaagct tggtaccgag ctcggatcca gtacccttca
     1201 ccatgtcgtc ctggataagg tggcatggac ccgccatggc gcggctctgg ggcttctgct
     1261 ggctggttgt gggcttctgg agggccgctt tcgcctgtcc cacgtcctgc aaatgcagtg
     1321 cctctcggat ctggtgcagc gacccttctc ctggcatcgt ggcatttccg agattggagc
     1381 ctaacagtgt agatcctgag aacatcaccg aaattttcat cgcaaaccag aaaaggttag
     1441 aaatcatcaa cgaagatgat gttgaagctt atgtgggact gagaaatctg acaattgtgg
     1501 attctggatt aaaatttgtg gctcataaag catttctgaa aaacagcaac ctgcagcaca
     1561 tcaattttac ccgaaacaaa ctgacgagtt tgtctaggaa acatttccgt caccttgact
     1621 tgtctgaact gatcctggtg ggcaatccat ttacatgctc ctgtgacatt atgtggatca
     1681 agactctcca agaggctaaa tccagtccag acactcagga tttgtactgc ctgaatgaaa
     1741 gcagcaagaa tattcccctg gcaaacctgc agatacccaa ttgtggtttg ccatctgcaa
     1801 atctggccgc acctaacctc actgtggagg aaggaaagtc tatcacatta tcctgtagtg
     1861 tggcaggtga tccggttcct aatatgtatt gggatgttgg taacctggtt tccaaacata
     1921 tgaatgaaac aagccacaca cagggctcct taaggataac taacatttca tccgatgaca
     1981 gtgggaagca gatctcttgt gtggcggaaa atcttgtagg agaagatcaa gattctgtca
     2041 acctcactgt gcattttgca ccaactatca catttctcga atctccaacc tcagaccacc
     2101 actggtgcat tccattcact gtgaaaggca accccaaacc agcgcttcag tggttctata
     2161 acggggcaat attgaatgag tccaaataca tctgtactaa aatacatgtt accaatcaca
     2221 cggagtacca cggctgcctc cagctggata atcccactca catgaacaat ggggactaca
     2281 ctctaatagc caagaatgag tatgggaagg atgagaaaca gatttctgct cacttcatgg
     2341 gctggcctgg aattgacgat ggtgcaaacc caaattatcc tgatgtaatt tatgaagatt
     2401 atggaactgc agcgaatgac atcggggaca ccacgaacag aagtaatgaa atcccttcca
     2461 cagacgtcac tgataaaacc ggtcgggaac atctctcggt ctatgctgtg gtggtgattg
     2521 cgtctgtggt gggattttgc cttttggtaa tgctgtttct gcttaagttg gcaagacact
     2581 ccaagtttgg catgaaaggc ccagcctccg ttatcagcaa tgatgatgac tctgccagcc
     2641 cactccatca catctccaat gggagtaaca ctccatcttc ttcggaaggt ggcccagatg
     2701 ctgtcattat tggaatgacc aagatccctg tcattgaaaa tccccagtac tttggcatca
     2761 ccaacagtca gctcaagcca gacacatttg ttcagcacat caagcgacat aacattgttc
     2821 tgaaaaggga gctaggcgaa ggagcctttg gaaaagtgtt cctagctgaa tgctataacc
     2881 tctgtcctga gcaggacaag atcttggtgg cagtgaagac cctgaaggat gccagtgaca
     2941 atgcacgcaa ggacttccac cgtgaggccg agctcctgac caacctccag catgagcaca
     3001 tcgtcaagtt ctatggcgtc tgcgtggagg gcgaccccct catcatggtc tttgagtaca
     3061 tgaagcatgg ggacctcaac aagttcctca gggcacacgg ccctgatgcc gtgctgatgg
     3121 ctgagggcaa cccgcccacg gaactgacgc agtcgcagat gctgcatata gcccagcaga
     3181 tcgccgcggg catggtctac ctggcgtccc agcacttcgt gcaccgcgat ttggccacca
     3241 ggaactgcct ggtcggggag aacttgctgg tgaaaatcgg ggactttggg atgtcccggg
     3301 acgtgtacag cactgactac tacagggtcg gtggccacac aatgctgccc attcgctgga
     3361 tgcctccaga gagcatcatg tacaggaaat tcacgacgga aagcgacgtc tggagcctgg
     3421 gggtcgtgtt gtgggagatt ttcacctatg gcaaacagcc ctggtaccag ctgtcaaaca
     3481 atgaggtgat agagtgtatc actcagggcc gagtcctgca gcgaccccgc acgtgccccc
     3541 aggaggtgta tgagctgatg ctggggtgct ggcagcgaga gccccacatg aggaagaaca
     3601 tcaagggcat ccataccctc cttcagaact tggccaaggc atctccggtc tacctggaca
     3661 ttctaggcaa gggtcaagac aattctgcag atatccagca cagtggcggc cgctcgagtc
     3721 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     3781 cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     3841 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3901 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3961 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4021 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4081 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4141 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4201 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4261 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4321 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4381 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4441 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4501 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4561 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4621 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4681 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4741 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4801 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4861 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4921 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4981 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5041 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5101 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5161 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5221 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5281 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5341 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5401 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5461 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5521 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5581 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5641 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5701 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     5761 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5821 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5881 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5941 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6001 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6061 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6901 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaa
//
