LOCUS       sequence-308800-        5361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST_CK1-alpha (human CK1-alpha as a fusion protein with 
            GST and His- tags) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5361)
  AUTHORS   Gavagan M, Fagnan E, Speltz EB, Zalatan JG
  TITLE     The Scaffold Protein Axin Promotes Signaling Specificity within the 
            Wnt Pathway by Suppressing Competing Kinase Reactions.
  JOURNAL   Cell Syst. 2020 Jun 24;10(6):515-525.e5. doi: 
            10.1016/j.cels.2020.05.002. Epub 2020 Jun 17.
  PUBMED    32553184
REFERENCE   2  (bases 1 to 5361)
  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: 
            "10.1016/j.cels.2020.05.002"; journalName: "Cell Syst"; date: 
            "2020-06-24- 24"; volume: "10"; issue: "6"; pages: "515-525.e5"
COMMENT     Sequence Label: pMG046 pETARA-CK1-alpha-His
FEATURES             Location/Qualifiers
     source          1..5361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(102..121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     312..333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        496..600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(601..669,670..1461)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(819..838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1632..2220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2121..2140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2374..2391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2406..2545
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2471..2490)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2631..2653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2647..2835)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="TKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHAD
                     ELYRSCLARFGDDGENL"
     primer_bind     3347..3366
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3347..3365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3397..3419
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3442..4092
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     4053..4075
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4129..4149
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5173..5190
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5208..5225
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5278..5296)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5292..5339
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
       61 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
      121 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
      181 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
      241 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
      301 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
      361 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
      421 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
      481 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      541 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      601 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      661 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      721 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      781 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      841 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      901 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      961 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1021 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1081 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1141 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1201 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1261 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1321 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1381 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1441 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1501 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1561 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1621 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1681 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1741 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1801 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1861 ctagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1921 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1981 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2041 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2101 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2161 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2221 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2281 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2341 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2401 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     2461 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     2521 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     2581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     2701 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     2761 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     2821 tttttcctgt ttggttactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     2881 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     2941 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3001 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     3061 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     3121 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     3181 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     3241 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     3301 ccgtggccag gacccaacgc tgcccgagat ctcgatcccg cgaaattaat acgactcact
     3361 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
     3421 atacatatga aacaagatct gatgtcccct atactaggtt attggaaaat taagggcctt
     3481 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
     3541 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
     3601 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
     3661 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
     3721 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
     3781 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     3841 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     3901 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     3961 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     4021 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     4081 gaccatcctc caactagtgg atctggtggt ggtggcggtt ccatgagcga gaatctttat
     4141 tttcagggat ccatggcgag tagcagcggc tccaaggctg aattcattgt cggagggaaa
     4201 tataaactgg tacggaagat cgggtctggc tccttcgggg acatctattt ggcgatcaac
     4261 atcaccaacg gcgaggaagt ggcagtgaag ctagaatctc agaaggccag gcatccccag
     4321 ttgctgtacg agagcaagct ctataagatt cttcaaggtg gggttggcat cccccacata
     4381 cggtggtatg gtcaggaaaa agactacaat gtactagtca tggatcttct gggacctagc
     4441 ctcgaagacc tcttcaattt ctgttcaaga aggttcacaa tgaaaactgt acttatgtta
     4501 gctgaccaga tgatcagtag aattgaatat gtgcatacaa agaattttat acacagagac
     4561 attaaaccag ataacttcct aatgggtatt gggcgtcact gtaataagtt attccttatt
     4621 gattttggtt tggccaaaaa gtacagagac aacaggacaa ggcaacacat accatacaga
     4681 gaagataaaa acctcactgg cactgcccga tatgctagca tcaatgcaca tcttggtatt
     4741 gagcagagtc gccgagatga catggaatca ttaggatatg ttttgatgta ttttaataga
     4801 accagcctgc catggcaagg gctaaaggct gcaacaaaga aacaaaaata tgaaaagatt
     4861 agtgaaaaga agatgtccac gcctgttgaa gttttatgta aggggtttcc tgcagaattt
     4921 gcgatgtact taaactattg tcgtgggcta cgctttgagg aagccccaga ttacatgtat
     4981 ctgaggcagc tattccgcat tcttttcagg accctgaacc atcaatatga ctacacattt
     5041 gattggacaa tgttaaagca gaaagcagca cagcaggcag cctcttccag tgggcagggt
     5101 cagcaggccc aaacccccac aggcaagcaa actgacaaat ccaagagtaa catgaaaggt
     5161 ttcagcggcc gccaccatca ccaccaccac taggggccgc actcgagcac caccaccacc
     5221 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     5281 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     5341 tgaaaggagg aactatatcc g
//
