LOCUS       sequence-110311-        5250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to create a cassette for homologous recombination in S. 
            cerevisiae; adds a C-terminal CBP-3xFLAG-TEV-ProtA triple tag and a 
            LEU2 selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5250)
  AUTHORS   Lin TY, Voronovsky A, Raabe M, Urlaub H, Sander B, Golas MM
  TITLE     Dual tagging as an approach to isolate endogenous chromatin 
            remodeling complexes from Saccharomyces cerevisiae.
  JOURNAL   Biochim Biophys Acta. 2015 Mar;1854(3):198-208. doi: 
            10.1016/j.bbapap.2014.11.009. Epub 2014 Dec 5.
  PUBMED    25486077
REFERENCE   2  (bases 1 to 5250)
  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.bbapap.2014.11.009"; journalName: "Biochim Biophys Acta";
            date: "2015-03"; volume: "1854"; issue: "3"; pages: "198-208"
COMMENT     Sequence Label: pGS1034
FEATURES             Location/Qualifiers
     source          1..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2256..2333
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2349..2414
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2442..2462
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2502..2675
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2676..2849
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     terminator      2905..3092
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2960..2984)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3167..3510
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(4856..4873)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4856..4872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4865..4887)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5081..5100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5200..5222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt ggttctggtt ctggtaaaag aagatggaag aagaatttta
     2281 ttgctgtttc tgctgctaat agatttaaga aaatttcttc ttctggtgct ttgtctgctt
     2341 ctggttctga ctacaaagac catgacggtg attataaaga tcatgacatc gattacaagg
     2401 atgacgatga caaggattat gatattccaa ctactgctag cgagaatttg tattttcagg
     2461 gtgagctcaa aaccgcggct cttgcgcaac acgatgaagc cgtggacaac aaattcaaca
     2521 aagaacaaca aaacgcgttc tatgagatct tacatttacc taacttaaac gaagaacaac
     2581 gaaacgcctt catccaaagt ttaaaagatg acccaagcca aagcgctaac cttttagcag
     2641 aagctaaaaa gctaaatgat gctcaggcgc cgaaagtaga caacaaattc aacaaagaac
     2701 aacaaaacgc gttctatgag atcttacatt tacctaactt aaacgaagaa caacgaaacg
     2761 ccttcatcca aagtttaaaa gatgacccaa gccaaagcgc taacctttta gcagaagcta
     2821 aaaagctaaa tggtgctcag gcgccgaaag tagacgcgaa ttccgcgggg aagtcaacct
     2881 gagtcgaccg ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     2941 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     3001 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat
     3061 gaggtcgctc ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata
     3121 tagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca
     3181 gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc
     3241 gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg
     3301 cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc
     3361 cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta
     3421 ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag
     3481 ttctcacatc acatccgaac ataaacaacc atgtctaaga atatcgttgt cctaccgggt
     3541 gatcacgtcg gtaaagaagt tactgacgaa gctattaagg tcttgaatgc cattgctgaa
     3601 gtccgtccag aaattaagtt caatttccaa catcacttga tcgggggtgc tgccatcgat
     3661 gccactggca ctcctttacc agatgaagct ctagaagcct ctaagaaagc cgatgctgtc
     3721 ttactaggtg ctgttggtgg tccaaaatgg ggtacgggcg cagttagacc agaacaaggt
     3781 ctattgaaga tcagaaagga attgggtcta tacgccaact tgagaccatg taactttgct
     3841 tctgattctt tactagatct ttctcctttg aagcctgaat atgcaaaggg taccgatttc
     3901 gtcgtcgtta gagaattggt tggtggtatc tactttggtg aaagaaaaga agatgaaggt
     3961 gacggagttg cttgggactc tgagaaatac agtgttcctg aagttcaaag aattacaaga
     4021 atggctgctt tcttggcatt gcaacaaaac ccaccattac caatctggtc tcttgacaag
     4081 gctaacgtgc ttgcctcttc cagattgtgg agaaagactg ttgaagaaac catcaagact
     4141 gagttcccac aattaactgt tcagcaccaa ttgatcgact ctgctgctat gattttggtt
     4201 aaatcaccaa ctaagctaaa cggtgttgtt attaccaaca acatgtttgg tgatattatc
     4261 tccgatgaag cctctgttat tccaggttct ttgggtttat taccttctgc atctctagct
     4321 tccctacctg acactaacaa ggcattcggt ttgtacgaac catgtcatgg ttctgcccca
     4381 gatttaccag caaacaaggt taacccaatt gctaccatct tatctgcagc tatgatgttg
     4441 aagttatcct tggatttggt tgaagaaggt agggctcttg aagaagctgt tagaaatgtc
     4501 ttggatgcag gtgtcagaac cggtgacctt ggtggttcta actctaccac tgaggttggc
     4561 gatgctatcg ccaaggctgt caaggaaatc ttggcttaaa gagtcttttg taacgacccc
     4621 gtctccacca acttggtatg cttgaaatct caaggccatt acacattcag ttatgtgaac
     4681 gaaaggtctt tatttaacgt agcataaact aaataataca ggttccggtt agcctgcaat
     4741 gtgttaaatc taaaggagca tacccaaaat gaactgaaga caaggaaatt tgcttgtcca
     4801 gatgtgattg gatatcagaa tctagatgca ttcgcgaggt accgagctcg aattcactgg
     4861 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4921 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     4981 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     5041 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     5101 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5161 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5221 ggttttcacc gtcatcaccg aaacgcgcga
//
