LOCUS       sequence-407173-        7991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TRKC receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7991)
  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 7991)
  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-TRKC-V5/HIS
FEATURES             Location/Qualifiers
     source          1..7991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     464..483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    770..891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(807..826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     861..880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(940..956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(940..956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(953..975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    964..980
                     /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(988..994,995..1012,1013..1018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1033..1054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1171..1188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1342..1930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1422..1441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2101..2892,2893..2961))
                     /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     2724..2743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2962..3066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3141..3160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3332..3711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3712..3915
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3866..3886
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3960..3979
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3960..3978
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6586..6627
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6637..6654
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6677..6694)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6683..6907
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6953..7381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7040..7059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7250..7271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7390..7410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7395..7724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7575..7710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7637..7656
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(7791..7991,1..594)
                     /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(7845..7864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
       61 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
      121 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
      181 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
      241 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
      301 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
      361 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
      421 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
      481 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
      541 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
      601 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
      661 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
      721 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
      781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
      841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
      901 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
      961 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1021 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1081 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1141 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1201 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1261 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1321 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1381 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1441 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1501 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1561 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1621 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1681 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1741 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1801 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1861 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1921 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1981 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2041 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2221 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2341 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2401 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2461 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2521 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2581 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2641 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2701 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2761 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2821 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2881 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2941 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3001 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3061 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     3121 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     3181 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     3241 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     3301 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     3361 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3421 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3481 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3541 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3601 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3661 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3721 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3781 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3841 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3901 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     3961 aatacgactc actataggga gacccaagct ggctagttaa gcttggtacc gagctcggat
     4021 ccagtaccct tcaccatgga tgtctctctt tgcccagcca agtgtagttt ctggcggatt
     4081 ttcttgctgg gaagcgtctg gctggactat gtgggctccg tgctggcttg ccctgcaaat
     4141 tgtgtctgca gcaagactga gatcaattgc cggcggccgg acgatgggaa cctcttcccc
     4201 ctcctggaag ggcaggattc agggaacagc aatgggaacg ccagtatcaa catcacggac
     4261 atctcaagga atatcacttc catacacata gagaactggc gcagtcttca cacgctcaac
     4321 gccgtggaca tggagctcta caccggactt caaaagctga ccatcaagaa ctcaggactt
     4381 cggagcattc agcccagagc ctttgccaag aacccccatt tgcgttatat aaacctgtca
     4441 agtaaccggc tcaccacact ctcgtggcag ctcttccaga cgctgagtct tcgggaattg
     4501 cagttggagc agaacttttt caactgcagc tgtgacatcc gctggatgca gctctggcag
     4561 gagcaggggg aggccaagct caacagccag aacctctact gcatcaacgc tgatggctcc
     4621 cagcttcctc tcttccgcat gaacatcagt cagtgtgacc ttcctgagat cagcgtgagc
     4681 cacgtcaacc tgaccgtacg agagggtgac aatgctgtta tcacttgcaa tggctctgga
     4741 tcaccccttc ctgatgtgga ctggatagtc actgggctgc agtccatcaa cactcaccag
     4801 accaatctga actggaccaa tgttcatgcc atcaacttga cgctggtgaa tgtgacgagt
     4861 gaggacaatg gcttcaccct gacgtgcatt gcagagaacg tggtgggcat gagcaatgcc
     4921 agtgttgccc tcactgtcta ctatccccca cgtgtggtga gcctggagga gcctgagctg
     4981 cgcctggagc actgcatcga gtttgtggtg cgtggcaacc ccccaccaac gctgcactgg
     5041 ctgcacaatg ggcagcctct gcgggagtcc aagatcatcc atgtggaata ctaccaagag
     5101 ggagagattt ccgagggctg cctgctcttc aacaagccca cccactacaa caatggcaac
     5161 tataccctca ttgccaaaaa cccactgggc acagccaacc agaccatcaa tggccacttc
     5221 ctcaaggagc cctttccaga gagcacggat aactttatct tgtttgacga agtgagtccc
     5281 acacctccta tcactgtgac ccacaaacca gaagaagaca cttttggggt atccatagca
     5341 gttggacttg ctgcttttgc ctgtgtcctg ttggtggttc tcttcgtcat gatcaacaaa
     5401 tatggtcgac ggtccaaatt tggaatgaag ggtcccgtgg ctgtcatcag tggtgaggag
     5461 gactcagcca gcccactgca ccacatcaac cacggcatca ccacgccctc gtcactggat
     5521 gccgggcccg acactgtggt cattggcatg actcgcatcc ctgtcattga gaacccccag
     5581 tacttccgtc agggacacaa ctgccacaag ccggacacgt atgtgcagca cattaagagg
     5641 agagacatcg tgctgaagcg agaactgggt gagggagcct ttggaaaggt cttcctggcc
     5701 gagtgctaca acctcagccc gaccaaggac aagatgcttg tggctgtgaa ggccctgaag
     5761 gatcccaccc tggctgcccg gaaggatttc cagagggagg ccgagctgct caccaacctg
     5821 cagcatgagc acattgtcaa gttctatgga gtgtgcggcg atggggaccc cctcatcatg
     5881 gtctttgaat acatgaagca tggagacctg aataagttcc tcagggccca tgggccagat
     5941 gcaatgatcc ttgtggatgg acagccacgc caggccaagg gtgagctggg gctctcccaa
     6001 atgctccaca ttgccagtca gatcgcctcg ggtatggtgt acctggcctc ccagcacttt
     6061 gtgcaccgag acctggccac caggaactgc ctggttggag cgaatctgct agtgaagatt
     6121 ggggacttcg gcatgtccag agatgtctac agcacggatt attacagggt gggaggacac
     6181 accatgctcc ccattcgctg gatgcctcct gaaagcatca tgtaccggaa gttcactaca
     6241 gagagtgatg tatggagctt cggggtgatc ctctgggaga tcttcaccta tggaaagcag
     6301 ccatggttcc aactctcaaa cacggaggtc attgagtgca ttacccaagg tcgtgttttg
     6361 gagcggcccc gagtctgccc caaagaggtg tacgatgtca tgctggggtg ctggcagagg
     6421 gaaccacagc agcggttgaa catcaaggag atctacaaaa tcctccatgc tttggggaag
     6481 gccaccccaa tctacctgga cattcttggc gcaaagggtc aagacaattc tgcagatatc
     6541 cagcacagtg gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct
     6601 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     6661 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6721 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6781 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6841 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     6901 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     6961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     7021 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     7081 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     7141 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     7201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7261 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     7321 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7381 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     7441 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     7501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     7561 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     7621 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     7681 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     7741 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     7801 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     7861 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     7921 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     7981 cagcgcggct a
//
