LOCUS       sequence-110117-        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to create a cassette for homologous recombination in S. 
            cerevisiae; adds a C-terminal 3xFLAG tag and a LEU2 selection 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  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 4692)
  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: pGS1044
FEATURES             Location/Qualifiers
     source          1..4692
                     /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..2321
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      2347..2534
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2402..2426)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2609..2952
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(4298..4315)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4298..4314)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4307..4329)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4523..4542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4642..4664
                     /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 tctgcttctg gttctgacta caaagaccat gacggtgatt
     2281 ataaagatca tgacatcgat tacaaggatg acgatgacaa gtaagtcgac cgccacttct
     2341 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     2401 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     2461 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
     2521 cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc
     2581 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
     2641 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
     2701 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
     2761 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
     2821 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
     2881 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
     2941 acataaacaa ccatgtctaa gaatatcgtt gtcctaccgg gtgatcacgt cggtaaagaa
     3001 gttactgacg aagctattaa ggtcttgaat gccattgctg aagtccgtcc agaaattaag
     3061 ttcaatttcc aacatcactt gatcgggggt gctgccatcg atgccactgg cactccttta
     3121 ccagatgaag ctctagaagc ctctaagaaa gccgatgctg tcttactagg tgctgttggt
     3181 ggtccaaaat ggggtacggg cgcagttaga ccagaacaag gtctattgaa gatcagaaag
     3241 gaattgggtc tatacgccaa cttgagacca tgtaactttg cttctgattc tttactagat
     3301 ctttctcctt tgaagcctga atatgcaaag ggtaccgatt tcgtcgtcgt tagagaattg
     3361 gttggtggta tctactttgg tgaaagaaaa gaagatgaag gtgacggagt tgcttgggac
     3421 tctgagaaat acagtgttcc tgaagttcaa agaattacaa gaatggctgc tttcttggca
     3481 ttgcaacaaa acccaccatt accaatctgg tctcttgaca aggctaacgt gcttgcctct
     3541 tccagattgt ggagaaagac tgttgaagaa accatcaaga ctgagttccc acaattaact
     3601 gttcagcacc aattgatcga ctctgctgct atgattttgg ttaaatcacc aactaagcta
     3661 aacggtgttg ttattaccaa caacatgttt ggtgatatta tctccgatga agcctctgtt
     3721 attccaggtt ctttgggttt attaccttct gcatctctag cttccctacc tgacactaac
     3781 aaggcattcg gtttgtacga accatgtcat ggttctgccc cagatttacc agcaaacaag
     3841 gttaacccaa ttgctaccat cttatctgca gctatgatgt tgaagttatc cttggatttg
     3901 gttgaagaag gtagggctct tgaagaagct gttagaaatg tcttggatgc aggtgtcaga
     3961 accggtgacc ttggtggttc taactctacc actgaggttg gcgatgctat cgccaaggct
     4021 gtcaaggaaa tcttggctta aagagtcttt tgtaacgacc ccgtctccac caacttggta
     4081 tgcttgaaat ctcaaggcca ttacacattc agttatgtga acgaaaggtc tttatttaac
     4141 gtagcataaa ctaaataata caggttccgg ttagcctgca atgtgttaaa tctaaaggag
     4201 catacccaaa atgaactgaa gacaaggaaa tttgcttgtc cagatgtgat tggatatcag
     4261 aatctagatg cattcgcgag gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     4321 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4381 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4441 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4501 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4561 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4621 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4681 cgaaacgcgc ga
//
