LOCUS       sequence-407210-        7346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human RYK receptor tyrosine kinase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7346)
  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 7346)
  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-RYK-V5/HIS
FEATURES             Location/Qualifiers
     source          1..7346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(445..549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(624..643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        815..1194
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1195..1398
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1349..1369
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1443..1462
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1443..1461
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3424..3465
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3475..3492
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3515..3532)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3521..3745
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3791..4219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3878..3897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4088..4109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4228..4248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4233..4562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4413..4548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4475..4494
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4629..5423
                     /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(4683..4702)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5293..5312
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5599..5720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5636..5655)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5690..5709
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5769..5785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5769..5785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5782..5804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5793..5809
                     /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(5817..5823,5824..5841,5842..5847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5862..5883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6000..6017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6171..6759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6251..6270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6930..7346,1..375,376..444))
                     /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"
ORIGIN
        1 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
       61 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      121 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      181 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      241 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      301 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      361 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      421 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      481 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      541 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
      601 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
      661 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
      721 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
      781 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
      841 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      901 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      961 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1021 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1081 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1141 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1201 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1261 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1321 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1381 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     1441 attaatacga ctcactatag ggagacccaa gctggctagt taagcttggt accgagctcg
     1501 gatccagtac ccttcaccat gcgtggggcg gcgcggctgg ggcggccggg ccggagttgc
     1561 ctcccggggg cccgcggcct gagggccccg ccgccgccgc cgctgctgct tctgcttgcg
     1621 ctgttgccgc tgctgcccgc gcctggcgct gccgccgccc ccgccccgcg gcccccggag
     1681 ctgcagtcgg cttccgcggg gcccagcgtg agtctctacc tgagcgagga cgaggtgcgc
     1741 cggctgatcg gtcttgatgc agaactttat tatgtgagaa atgaccttat tagtcactac
     1801 gctctatcct ttagtctgtt agtacccagt gagacaaatt tcctgcactt cacctggcat
     1861 gcgaagtcca aggttgaata taagctggga ttccaagtgg acaatgtttt ggcaatggat
     1921 atgccccagg tcaacatttc tgttcagggg gaagttccac gcactttatc agtgtttcgg
     1981 gtagagcttt cctgtactgg caaagtagat tctgaagtta tgatactaat gcagctcaac
     2041 ttgacagtaa attcttcaaa aaattttacc gtcttaaatt ttaaacgaag gaaaatgtgc
     2101 tacaaaaaac ttgaagaagt aaaaacttca gccttggaca aaaacactag cagaactatt
     2161 tatgatcctg tacatgcagc tccaaccact tctacgcgtg tgttttatat tagtgtaggg
     2221 gtttgttgtg cagtaatatt tctcgtagca ataatattag ctgttttgca ccttcatagt
     2281 atgaaaagga ttgaactgga tgacagcatt agtgccagca gtagttccca agggctgtct
     2341 cagccatcca cccagacgac tcagtatctg agagcagaca cgcccaacaa tgcaactcct
     2401 atcaccagct ccttaggtta tcctaccttg cggatagaga agaacgactt gagaagtgtc
     2461 actcttttgg aggccaaagg caaggtgaag gatatagcaa tatccagaga gaggataact
     2521 ctaaaagatg tactccaaga aggtactttt gggcgtattt tccatgggat tttaatagat
     2581 gaaaaagatc caaataaaga aaaacaagca tttgtcaaaa cagttaaaga tcaagcttct
     2641 gaaattcagg tgacaatgat gctcactgaa agttgtaagc tgcgaggtct tcatcacaga
     2701 aatcttcttc ctattactca tgtgtgtata gaagaaggag aaaagcccat ggtgatattg
     2761 ccttacatga attgggggaa tcttaaattg tttttacgac agtgcaagtt agtagaggcc
     2821 aataatccac aggcaatttc tcagcaagac ctggtacaca tggctattca gattgcctgt
     2881 ggaatgagct acctggccag aagggaagtc atccacaaag acctggctgc caggaactgt
     2941 gtcattgatg acacacttca agttaagatc acagacaatg ccctctccag agacttgttc
     3001 cccatggact atcactgtct gggggacaat gaaaacaggc cagttcgttg gatggctctt
     3061 gaaagtctgg ttaataacga gttctctagc gctagtgatg tgtgggcctt tggagtgacg
     3121 ctgtgggaac tcatgactct gggccagact ccctacgtgg acattgaccc cttcgagatg
     3181 gccgcatacc tgaaagatgg ttaccgaata gcccagccaa tcaactgtcc tgatgaatta
     3241 tttgctgtga tggcctgttg ctgggcctta gatccagagg agaggcccaa gtttcagcag
     3301 ctggtacagt gcctaacaga gtttcatgca gccctggggg cctacgtcgg gaagggtcaa
     3361 gacaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
     3421 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat
     3481 caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     3541 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3601 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3661 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3721 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3781 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3841 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3901 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3961 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4021 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4081 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4141 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4201 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4381 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4441 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4501 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4561 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4621 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4681 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4741 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4801 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4861 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4921 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4981 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5041 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5101 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5161 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5221 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5281 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5341 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5401 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     5461 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5521 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5581 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5641 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5701 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7321 acatgatccc ccatgttgtg caaaaa
//
