LOCUS       Exported                4917 bp ds-DNA     circular SYN 14-MAY-2024
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            TagRFp657 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4917)
  AUTHORS   Lee S, Lim WA, Thorn KS
  TITLE     Improved Blue, Green, and Red Fluorescent Protein Tagging Vectors 
            for S. cerevisiae.
  JOURNAL   PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902.
            Print 2013.
  PUBMED    23844123
REFERENCE   2  (bases 1 to 4917)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4917
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     125..143
                     /locus_tag="pBRforEco"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(181..203)
                     /locus_tag="pGEX 3'"
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     303..322
                     /locus_tag="pRS-marker"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     promoter        403..421
                     /locus_tag="SP6 promoter"
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     403..420
                     /locus_tag="SP6"
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    491..1210
                     /locus_tag="ymTq2"
                     /label=ymTq2
     terminator      1251..1438
                     /gene="S. cerevisiae ADH1"
                     /locus_tag="ADH1 terminator"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1306..1330)
                     /locus_tag="pHybLex-R"
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1513..2869
                     /locus_tag="kanMX"
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        1513..1856
                     /locus_tag="TEF promoter"
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1857..2666
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /locus_tag="KanR"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1924..1943)
                     /locus_tag="Kan-R"
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2672..2869
                     /locus_tag="TEF terminator"
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2973..2992)
                     /locus_tag="T7"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2974..2992)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3079..3096)
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3250..3838)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3330..3349)
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4009..4869)
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4632..4651
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4870..4917,1..57))
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
      181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      301 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
      361 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
      421 aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cggatcggtg acggtgctgg
      481 tttaattaac atggttagta aaggtgaaga attgttcact ggtgtagtgc caattttagt
      541 agaattagat ggtgacgtta acggtcataa gttctctgta agcggtgaag gtgaaggtga
      601 cgcaacatac ggtaaattaa ctttgaaatt catttgtact acaggtaaat taccagttcc
      661 atggccaact ttggttacta ctttgtcttg gggtgttcaa tgtttcgcta gatacccaga
      721 tcatatgaag caacatgatt tctttaagtc tgctatgcca gaaggttacg ttcaagaaag
      781 aactatcttc ttcaaggatg atggtaacta taagactaga gctgaagtta agttcgaagg
      841 agatacttta gttaatagaa ttgaattgaa gggtattgat tttaaggaag atggtaatat
      901 tttgggtcat aagttggaat ataattactt ctctgataac gtttacatta ctgctgataa
      961 gcaaaagaac ggtattaagg cgaacttcaa gattagacat aacattgaag atggtggtgt
     1021 tcaattagct gatcattacc aacaaaacac tccaattggt gacggtccag ttttgttgcc
     1081 agataaccat tacttgtcta ctcaatctaa gttgtctaag gatccaaacg aaaagagaga
     1141 tcatatggtt ttgttggaat tcgttactgc tgctgggatc actttaggta tggatgaatt
     1201 gtataaataa gcggccgcct cgaggtcgac ggcgcgccac ttctaaataa gcgaatttct
     1261 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
     1321 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
     1381 tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca
     1441 gatccgctag ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt
     1501 cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc
     1561 tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca
     1621 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
     1681 cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
     1741 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
     1801 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg
     1861 gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc caacatggat gctgatttat
     1921 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     1981 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2041 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     2101 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggca
     2161 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     2221 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     2281 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     2341 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     2401 agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2461 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     2521 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2581 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2641 atttgatgct cgatgagttt ttctaatcag tactgacaat aaaaagattc ttgttttcaa
     2701 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
     2761 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     2821 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg
     2881 atactaacgc cgccatccag tttaaacgag ctcgaattca tcgatgatat cagatccact
     2941 agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     3001 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3421 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     3481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3661 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtct
//
