LOCUS       sequence-388242-        8424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of  FLAG-tagged TAOK2 / PSK1 alpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8424)
  AUTHORS   Koo CY, Giacomini C, Reyes-Corral M, Olmos Y, Tavares IA, Marson CM,
            Linardopoulos S, Tutt AN, Morris JDH
  TITLE     Targeting TAO Kinases Using a New Inhibitor Compound Delays Mitosis 
            and Induces Mitotic Cell Death in Centrosome Amplified Breast Cancer
            Cells.
  JOURNAL   Mol Cancer Ther. 2017 Nov;16(11):2410-2421. doi: 
            10.1158/1535-7163.MCT-17-0077. Epub 2017 Aug 22.
  PUBMED    28830982
REFERENCE   2  (bases 1 to 8424)
  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"; doi: 
            "10.1158/1535-7163.MCT-17-0077"; journalName: "Mol Cancer Ther"; 
            date: "2017-11"; volume: "16"; issue: "11"; pages: "2410-2421"
COMMENT     Sequence Label: CMV-FLAG-TAOK2
FEATURES             Location/Qualifiers
     source          1..8424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4765..5387
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4964..4983)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5411..5431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5416..5745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5596..5731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5783..5802)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5784..5802)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5816..5832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5816..5832)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5829..5851)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5840..5856
                     /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(5864..5870,5871..5888,5889..5894))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5909..5930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6047..6064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6218..6806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6298..6317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6977..7768,7769..7837))
                     /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     7600..7619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7838..7942)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7969..8397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8056..8075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8266..8287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tatgccagct gggggccggg ccgggagcct gaaggaccca gatgtggctg agctcttctt
     1021 caaggatgac ccagaaaagc tcttctctga cctccgggaa attggccatg gcagctttgg
     1081 agccgtatac tttgcccggg atgtccggaa tagtgaggtg gtggccatca agaagatgtc
     1141 ctacagtggg aagcagtcca atgagaaatg gcaagacatc atcaaggagg tgcggttctt
     1201 acagaagctc cggcatccca acaccattca gtaccggggc tgttacctga gggagcacac
     1261 ggcttggctg gtaatggagt attgcctggg ctcagcttct gaccttctag aagtgcacaa
     1321 gaaacccctt caggaggtag agatcgcagc tgtgacccac ggggcgcttc agggcctggc
     1381 atatctgcac tcccacaaca tgatccatag ggatgtgaag gctggaaata tcctgctgtc
     1441 agagccaggg ttagtgaagc taggggactt tggttctgcg tccatcatgg cacctgccaa
     1501 ctccttcgtg ggcaccccat actggatggc acccgaggtg atcctggcca tggatgaggg
     1561 gcagtacgat ggcaaagtgg acgtctggtc cttggggata acctgcatcg agctggctga
     1621 acggaaacca ccgctcttta acatgaatgc gatgagtgcc ttataccaca ttgcacagaa
     1681 cgaatccccc gtgctccagt caggacactg gtctgagtac ttccggaatt ttgtcgactc
     1741 ctgtcttcag aaaatccctc aagacagacc aacctcagag gttctcctga agcaccgctt
     1801 tgtgctccgg gagcggccac ccacagtcat catggacctg atccagagga ccaaggatgc
     1861 cgtgcgggag ctggacaacc tgcagtaccg caagatgaag aagatcctgt tccaagaggc
     1921 acccaacggc cctggtgccg aggccccaga ggaggaagag gaggccgagc cctacatgca
     1981 ccgggccggg actctgacca gcctcgagag tagccactca gtgcccagca tgtccatcag
     2041 cgcctccagc cagagcagct ccgtcaacag cctagcagat gcctcagaca acgaggaaga
     2101 ggaggaggag gaggaggaag aggaggagga ggaagaaggc cctgaagccc gggagatggc
     2161 catgatgcag gagggggagc acacagtcac ctctcacagc tccattatcc accggctgcc
     2221 gggctctgac aacctatatg atgaccccta ccagccagag ataaccccca gccctctcca
     2281 gccgcctgca gccccagctc ccacttccac cacctcttcc gcccgccgcc gggcctactg
     2341 ccgtaaccga gaccactttg ccaccatccg aaccgcctcc ctggtcagcc gtcagatcca
     2401 ggagcatgag caggactctg cgctgcggga gcagctgagc ggctataagc ggatgcgacg
     2461 acagcaccag aagcagctgc tggccctgga gtcacggctg aggggtgaac gggaggagca
     2521 cagtgcacgg ctgcagcggg agcttgaggc gcagcgggct ggctttgggg cagaggcaga
     2581 aaagctggcc cggcggcacc aggccatagg tgagaaggag gcacgagctg cccaggccga
     2641 ggagcggaag ttccagcagc acatccttgg gcagcagaag aaggagctgg ctgccctgct
     2701 ggaggcacag aagcggacct acaaacttcg caaggaacag ctgaaggagg agctccagga
     2761 gaaccccagc actcccaagc gggagaaggc cgagtggctg ctgcggcaga aggagcagct
     2821 ccagcagtgc caggcggagg aggaagcagg gctgctgcgg cggcagcgcc agtactttga
     2881 gctgcagtgt cgccagtaca agcgcaagat gttgctggct cggcacagcc tggaccagga
     2941 cctgctgcgg gaggacctga acaagaagca gacccagaag gacttggagt gtgcactgct
     3001 gcttcggcag cacgaggcca cgcgggagct ggagctgcgg cagctccagg ccgtgcagcg
     3061 cacgcgggct gagctcaccc gcctgcagca ccagacggag ctgggcaacc agctggagta
     3121 caacaagcgg cgtgagcaag agttgcggca gaagcatgcg gcccaggttc gccagcagcc
     3181 caagagcctc aaagtacgtg caggccagcg ccccccgggc cttccactcc ccattcctgg
     3241 ggctctgggc ccacccaaca caggcacccc tatagaacag cagccctgct cacctggcca
     3301 ggaggcagtc ctggaccaaa gaatgcttgg cgaggaggag gaagcagttg gagagagaag
     3361 gattctggga aaggaagggg ccactttgga gcccaagcag cagaggattc tgggggaaga
     3421 atcaggagcc cctagtccca gtccacaaaa acatgggagc ctggttgatg aggaagtttg
     3481 gggtctgcct gaggagatag aggagcttag ggtgccctcc cttgtacccc aggagaggag
     3541 cattgttggc caggaggagg ctgggacgtg gagcttgtgg gggaaggagg atgagagtct
     3601 tctggatgag gagtttgagc ttggctgggt ccagggccca gcactgactc ccgtccctga
     3661 ggaggaggaa gaagaggaag agggggctcc gattgggacc cctagggatc ctggagatgg
     3721 ttgtccttcc cccgacatcc ctcctgaacc ccctccaaca cacctgaggc cctgccctgc
     3781 cagccagctc cctggactcc tgtcccatgg cctcctggcc ggcctctcct ttgcagtggg
     3841 gtcctcctct ggcctcctgc ccctcctgct gctgctgctg cttccattgc tggcagccca
     3901 gggtgggggt ggcctgcagg cagcgctgct ggcccttgag gtggggctgg tgggtctggg
     3961 ggcctcctac ctgctccttt gtacagccct gcacctgccc tccagtcttt tcctactcct
     4021 ggcccagggt accgcactgg gggccgtcct gggcctgagc tggcgccgag gcctcatggg
     4081 tgttcccctg ggccttggag ctgcctggct cttagcttgg ccaggcctag ctctacctct
     4141 ggtggctatg gcagcggggg gcagatgggt gcggcagcag ggcccccggg tgcgccgggg
     4201 catatctcga ctctggttgc gggttctgct gcgcctgtca cccatggcct tccgggccct
     4261 gcagggctgt ggggctgtgg gggaccgggg tctgtttgca ctgtacccca aaaccaacaa
     4321 ggatggcttc cgcagccgcc tgcccgtccc tgggccccgg cggcgtaatc cccgcaccac
     4381 ccaacaccca ttagctctgt tggcaagggt ctgggtcctg tgcaagggct ggaactggcg
     4441 tctggcacgg gccagccagg gtttagcatc ccacttgccc ccgtgggcca tccacacact
     4501 ggccagctgg ggcctgcttc ggggtgaacg gcccacccga atcccccggc tactaccacg
     4561 cagccagcgc cagctagggc cccctgcctc ccaccagcca ctgccaggga ctctagccgg
     4621 gcggaggtca cgcacccgcc agtcccgggc cctgcccccc tggaggtagc tgactccagc
     4681 ccttccagcc caaatctaga gcattgagta gatagaattc atcgatagat ctgatatcgg
     4741 taccagtcga ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     4801 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     4861 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     4921 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     4981 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     5041 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     5101 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     5161 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     5221 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     5281 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     5341 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     5401 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5461 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5521 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5581 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5641 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5701 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa
     5761 ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat
     5821 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5881 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5941 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6001 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6061 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6121 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6181 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6241 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6301 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6361 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6421 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6481 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6541 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6601 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6661 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6721 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6781 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6841 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6901 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6961 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7021 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7081 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7141 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7201 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7261 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7321 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7381 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7441 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7501 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7561 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7621 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7681 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7741 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7801 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7861 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7921 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     7981 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     8041 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     8101 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     8161 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     8221 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     8281 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     8341 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     8401 acaaaatatt aacgcttaca attt
//
