LOCUS       sequence-149772-        3251 bp    DNA     circular SYN 14-APR-2025
DEFINITION  N-terminal tagging of human EZH2 with 3xFLAG_Twin_Strep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3251)
  AUTHORS   Dalvai M, Loehr J, Jacquet K, Huard CC, Roques C, Herst P, Cote J, 
            Doyon Y
  TITLE     A Scalable Genome-Editing-Based Approach for Mapping Multiprotein 
            Complexes in Human Cells.
  JOURNAL   Cell Rep. 2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 
            10.1016/j.celrep.2015.09.009.
  PUBMED    26456817
REFERENCE   2  (bases 1 to 3251)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep. 
            2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 
            10.1016/j.celrep.2015.09.009."
COMMENT     Sequence Label: EZH2_N_FS_v2_Donor
FEATURES             Location/Qualifiers
     source          1..3251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     regulatory      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             617..682
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(692..697,698..721,722..757,758..781)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(1261..1278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1432..2020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1512..1531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2191..2982,2983..3051))
                     /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     2814..2833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3052..3156)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3224..3242
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatggt accattatgc cttgctactg taacagctaa tgatgaaaaa caggatctgt
      241 ctttatattt tcttccctcc acaaatgtgg atctcataga gttgaaaact aggttgtgat
      301 atagtatagt atacctaatt cctgtaatgg gatcatgttc ctataatatg gccgcaattt
      361 agtgtagaat ttttgtaaat aaaagtgtat tttaagttta acttaaactt tcaatgaagt
      421 gttttaagga tttaaccatg cagcacaaat gagcaccttt ctgtaaatgc caacagtgta
      481 atatgtgtca tttcttcact gattgttagt ttgctgcgga ttaaaacaca ggtgatcata
      541 ttcaggctgg ttagattagt gattttaata tgaaaccatt gcttttagaa taatcgccac
      601 catgggatcc gccggcgact acaaggacca cgacggcgat tataaggatc acgacatcga
      661 ctacaaagac gacgatgaca agggcgccag cagcgcctgg tcccaccctc agtttgagaa
      721 gggcggaggc tctggcggcg gaagcggagg atctgcttgg agccaccccc agttcgaaaa
      781 gggcgccggc cagactggga agaaatctga gaagggacca gtttgttggc ggaagcgtgt
      841 aaaatcagag tacatgcgac tgagacagct caagaggttc agacgagctg atgaagtaaa
      901 ggtataattt tatcttttgt gaaaatgaat atacaattaa ggataggttt tttcccctcc
      961 taagttcaat aagtttttat ttaaaataat tctagccaag tttgtattta caataagatg
     1021 agtgaatact tggaagctta ggttcttgat ctgtcttaac attataccat gcacattaat
     1081 cagtctttgt ttagtatact tttatttgca tatgcagctg cagaaatgcc tccctaaata
     1141 taaagggatg catttcatga ttttatgttt ctaaagtcat tgtgactcga gaattctgca
     1201 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1261 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1321 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1861 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3181 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3241 gccctttcgt c
//
