LOCUS       sequence-469590-        8144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the F866Y kinase defective mutant variant of human 
            DDR1 that might be associated with lung Adenocarcinoma.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  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 8144)
  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-DDR1-F866Y-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3641..3682
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3692..3709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3732..3749)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3738..3962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4008..4436
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4095..4114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4305..4326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4445..4465)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4450..4779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4630..4765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4692..4711
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4846..5640
                     /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(4900..4919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5510..5529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5816..5937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5853..5872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5907..5926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5986..6002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5986..6002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5999..6021)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6010..6026
                     /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(6034..6040,6041..6058,6059..6064))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6079..6100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6217..6234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6388..6976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6468..6487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7147..7938,7939..8007))
                     /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     7770..7789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8008..8112)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagtt
      901 aagcttggta ccgagctcgg atccagtacc cttcaccatg ggaccagagg ccctgtcatc
      961 tttactgctg ctgctcttgg tggcaagtgg agatgctgac atgaagggac attttgatcc
     1021 tgccaagtgc cgctatgccc tgggcatgca ggaccggacc atcccagaca gtgacatctc
     1081 tgcttccagc tcctggtcag attccactgc cgcccgccac agcaggttgg agagcagtga
     1141 cggggatggg gcctggtgcc ccgcagggtc ggtgtttccc aaggaggagg agtacttgca
     1201 ggtggatcta caacgactgc acctggtggc tctggtgggc acccagggac ggcatgccgg
     1261 gggcctgggc aaggagttct cccggagcta ccggctgcgt tactcccggg atggtcgccg
     1321 ctggatgggc tggaaggacc gctggggtca ggaggtgatc tcaggcaatg aggaccctga
     1381 gggagtggtg ctgaaggacc ttgggccccc catggttgcc cgactggttc gcttctaccc
     1441 ccgggctgac cgggtcatga gcgtctgtct gcgggtagag ctctatggct gcctctggag
     1501 ggatggactc ctgtcttaca ctgcccctgt ggggcagaca atgtatttat ctgaggccgt
     1561 gtacctcaac gactccacct atgacggaca taccgtgggc ggactgcagt atgggggtct
     1621 gggccagctg gcagatggtg tggtggggct ggatgacttt aggaagagtc aggagctgcg
     1681 ggtctggcca ggctatgact atgtgggatg gagcaaccac agcttctcca gtggctatgt
     1741 ggagatggag tttgagtttg accggctgag ggccttccag gctatgcagg tccactgtaa
     1801 caacatgcac acgctgggag cccgtctgcc tggcggggtg gaatgtcgct tccggcgtgg
     1861 ccctgccatg gcctgggagg gggagcccat gcgccacaac ctagggggca acctggggga
     1921 ccccagagcc cgggctgtct cagtgcccct tggcggccgt gtggctcgct ttctgcagtg
     1981 ccgcttcctc tttgcggggc cctggttact cttcagcgaa atctccttca tctctgatgt
     2041 ggtgaacaat tcctctccgg cactgggagg caccttcccg ccagccccct ggtggccgcc
     2101 tggcccacct cccaccaact tcagcagctt ggagctggag cccagaggcc agcagcccgt
     2161 ggccaaggcc gaggggagcc cgaccgccat cctcattggc tgcctggtgg ccatcatcct
     2221 gctcctgctg ctcatcattg ccctcatgct ctggcggctg cactggcgca ggctcctcag
     2281 caaggctgaa cggagggtgt tggaagagga gctgacggtt cacctctctg tccctgggga
     2341 cactatcctc atcaacaacc gcccaggtcc tagagagcca cccccgtacc aggagccccg
     2401 gcctcgtggg aatccgcccc actccgctcc ctgtgtcccc aatggctctg cctacagtgg
     2461 ggactatatg gagcctgaga agccaggcgc cccgcttctg cccccacctc cccagaacag
     2521 cgtcccccat tatgccgagg ctgacattgt taccctgcag ggcgtcaccg ggggcaacac
     2581 ctatgctgtg cctgcactgc ccccaggggc agtcggggat gggcccccca gagtggattt
     2641 ccctcgatct cgactccgct tcaaggagaa gcttggcgag ggccagtttg gggaggtgca
     2701 cctgtgtgag gtcgacagcc ctcaagatct ggttagtctt gatttccccc ttaatgtgcg
     2761 taagggacac cctttgctgg tagctgtcaa gatcttacgg ccagatgcca ccaagaatgc
     2821 caggaatgat ttcctgaaag aggtgaagat catgtcgagg ctcaaggacc caaacatcat
     2881 tcggctgctg ggcgtgtgtg tgcaggacga ccccctctgc atgattactg actacatgga
     2941 gaacggcgac ctcaaccagt tcctcagtgc ccaccagctg gaggacaagg cagccgaggg
     3001 ggcccctggg gacgggcagg ctgcgcaggg gcccaccatc agctacccaa tgctgctgca
     3061 tgtggcagcc cagatcgcct ccggcatgcg ctatctggcc acactcaact ttgtacatcg
     3121 ggacctggcc acgcggaact gcctagttgg ggaaaatttc accatcaaaa tcgcagactt
     3181 tggcatgagc cggaacctct atgctgggga ctattaccgt gtgcagggcc gggcagtgct
     3241 gcccatccgc tggatggcct gggagtgcat cctcatgggg aagttcacga ctgcgagtga
     3301 cgtgtgggcc tttggtgtga ccctgtggga ggtgctgatg ctctgtaggg cccagccctt
     3361 tgggcagctc accgacgagc aggtcatcga gaacgcgggg gagtatttcc gggaccaggg
     3421 ccggcaggtg tacctgtccc ggccgcctgc ctgcccgcag ggcctatatg agctgatgct
     3481 tcggtgctgg agccgggagt ctgagcagcg accacccttt tcccagctgc atcggttcct
     3541 ggcagaggat gcactcaaca cggtgaaaaa gggtcaagac aattctgcag atatccagca
     3601 cagtggcggc cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
     3661 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     3721 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3781 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3841 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3901 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3961 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4021 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4081 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4141 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4201 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4261 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4321 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4381 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4441 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4501 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4561 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4621 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4681 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4741 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4801 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4861 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4921 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4981 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5041 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5101 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5161 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5221 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5281 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5341 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5401 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5461 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5521 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5581 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5641 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5701 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5761 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5821 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5881 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5941 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6001 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6061 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6121 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6181 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6241 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6301 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6361 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6421 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6481 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6541 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6601 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6661 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6721 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6781 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6841 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6901 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6961 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7021 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7081 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7141 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7201 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7261 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7321 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7381 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7441 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7501 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7561 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7621 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7681 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7741 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7801 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7861 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7921 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7981 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8041 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8101 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcg
//
