LOCUS       sequence-394494-        8078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human DDR2 receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8078)
  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 8078)
  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-DDR2-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..52)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        224..603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        604..807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     758..778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     852..871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        852..870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1824..1832
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3565..3606
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3616..3633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3656..3673)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3662..3886
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3932..4360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4019..4038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4229..4250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4369..4389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4374..4703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4554..4689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4616..4635
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4770..5564
                     /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(4824..4843)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5434..5453
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5740..5861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5777..5796)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5831..5850
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5910..5926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5910..5926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5923..5945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5934..5950
                     /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(5958..5964,5965..5982,5983..5988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6003..6024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6141..6158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6312..6900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6392..6411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7071..7862,7863..7931))
                     /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     7694..7713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7932..8036)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
       61 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
      121 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
      181 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
      241 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      301 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      361 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      421 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      481 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      541 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      601 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      661 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      721 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      781 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
      841 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagtt aagcttggta
      901 ccgagctcgg atccagtacc cttcaccatg atcctgattc ccagaatgct cttggtgctg
      961 ttcctgctgc tgcctatctt gagttctgca aaagctcagg ttaatccagc tatatgccgc
     1021 tatcctctgg gcatgtcagg aggccagatt ccagatgagg acatcacagc ttccagtcag
     1081 tggtcagagt ccacagctgc caaatatgga aggctggact cagaagaagg ggatggagcc
     1141 tggtgccctg agattccagt ggaacctgat gacctgaagg agtttctgca gattgacttg
     1201 cacaccctcc attttatcac tctggtgggg acccaggggc gccatgcagg aggtcatggc
     1261 atcgagtttg cccccatgta caagatcaat tacagtcggg atggcactcg ctggatctct
     1321 tggcggaacc gtcatgggaa acaggtgctg gatggaaata gtaaccccta tgacattttc
     1381 ctaaaggact tggagccgcc cattgtagcc agatttgtcc ggttcattcc agtcaccgac
     1441 cactccatga atgtgtgtat gagagtggag ctttacggct gtgtctggct agatggcttg
     1501 gtgtcttaca atgctccagc tgggcagcag tttgtactcc ctggaggttc catcatttat
     1561 ctgaatgatt ctgtctatga tggagctgtt ggatacagca tgacagaagg gctaggccaa
     1621 ttgaccgatg gtgtgtctgg cctggacgat ttcacccaga cccatgaata ccacgtgtgg
     1681 cccggctatg actatgtggg ctggcggaac gagagtgcca ccaatggcta cattgagatc
     1741 atgtttgaat ttgaccgcat caggaatttc actaccatga aggtccactg caacaacatg
     1801 tttgctaaag gtgtgaagat ctttaaggag gtacagtgct acttccgctc tgaagccagt
     1861 gagtgggaac ctaatgccat ttccttcccc cttgtcctgg atgacgtcaa ccccagtgct
     1921 cggtttgtca cggtgcctct ccaccaccga atggccagtg ccatcaagtg tcaataccat
     1981 tttgcagata cctggatgat gttcagtgag atcaccttcc aatcagatgc tgcaatgtac
     2041 aacaactctg aagccctgcc cacctctcct atggcaccca caacctatga tccaatgctt
     2101 aaagttgatg acagcaacac tcggatcctg attggctgct tggtggccat catctttatc
     2161 ctcctggcca tcattgtcat catcctctgg aggcagttct ggcagaaaat gctggagaag
     2221 gcttctcgga ggatgctgga tgatgaaatg acagtcagcc tttccctgcc aagtgattct
     2281 agcatgttca acaataaccg ctcctcatca cctagtgaac aagggtccaa ctcgacttac
     2341 gatcgcatct ttccccttcg ccctgactac caggagccat ccaggctgat acgaaaactc
     2401 ccagaatttg ctccagggga ggaggagtca ggctgcagcg gtgttgtgaa gccagtccag
     2461 cccagtggcc ctgagggggt gccccactat gcagaggctg acatagtgaa cctccaagga
     2521 gtgacaggag gcaacacata ctcagtgcct gccgtcacca tggacctgct ctcaggaaaa
     2581 gatgtggctg tggaggagtt ccccaggaaa ctcctaactt tcaaagagaa gctgggagaa
     2641 ggacagtttg gggaggttca tctctgtgaa gtggagggaa tggaaaaatt caaagacaaa
     2701 gattttgccc tagatgtcag tgccaaccag cctgtcctgg tggctgtgaa aatgctccga
     2761 gcagatgcca acaagaatgc caggaatgat tttcttaagg agataaagat catgtctcgg
     2821 ctcaaggacc caaacatcat ccatctatta gctgtgtgta tcactgatga ccctctctgt
     2881 atgatcactg aatacatgga gaatggagat ctcaatcagt ttctttcccg ccacgagccc
     2941 cctaattctt cctccagcga tgtacgcact gtcagttaca ccaatctgaa gtttatggct
     3001 acccaaattg cctctggcat gaagtacctt tcctctctta attttgttca ccgagatctg
     3061 gccacacgaa actgtttagt gggtaagaac tacacaatca agatagctga ctttggaatg
     3121 agcaggaacc tgtacagtgg tgactattac cggatccagg gccgggcagt gctccctatc
     3181 cgctggatgt cttgggagag tatcttgctg ggcaagttca ctacagcaag tgatgtgtgg
     3241 gcctttgggg ttactttgtg ggagactttc accttttgtc aagaacagcc ctattcccag
     3301 ctgtcagatg aacaggttat tgagaatact ggagagttct tccgagacca agggaggcag
     3361 acttacctcc ctcaaccagc catttgtcct gactctgtgt ataagctgat gctcagctgc
     3421 tggagaagag atacgaagaa ccgtccctca ttccaagaaa tccaccttct gctccttcaa
     3481 caaggcgacg aaaagggtca agacaattct gcagatatcc agcacagtgg cggccgctcg
     3541 agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat
     3601 tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg
     3661 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3721 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3781 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3841 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3901 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3961 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4021 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4081 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4141 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4201 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4261 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4321 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4381 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4501 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4561 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4621 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4681 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4741 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4801 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4861 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4921 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4981 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5041 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5101 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5161 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5221 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5281 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5341 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5401 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5461 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5521 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5581 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5641 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5701 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5761 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5821 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5881 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5941 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6001 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6061 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6121 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6181 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6241 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6301 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6361 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6421 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6481 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6541 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6601 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6661 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6721 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6781 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6841 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6901 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6961 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7021 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7081 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7141 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7201 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7261 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7321 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7381 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7441 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7501 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7561 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7621 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7681 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7741 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7801 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7861 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7921 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7981 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8041 tttccccgaa aagtgccacc tgacgtcgac ggatcggg
//
