LOCUS       Exported                4907 bp ds-DNA     circular SYN 15-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 4907)
  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 4907)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4907
                     /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    490..1197
                     /locus_tag="yeast optimized mneongreen"
                     /label=yeast optimized mneongreen
     terminator      1241..1428
                     /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(1296..1320)
                     /locus_tag="pHybLex-R"
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1503..2859
                     /locus_tag="kanMX"
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        1503..1846
                     /locus_tag="TEF promoter"
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1847..2656
                     /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(1914..1933)
                     /locus_tag="Kan-R"
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2662..2859
                     /locus_tag="TEF terminator"
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2963..2982)
                     /locus_tag="T7"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2964..2982)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3069..3086)
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3240..3828)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3320..3339)
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3999..4859)
                     /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     4622..4641
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4860..4907,1..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 tttaattaaa tggtctctaa gggtgaagaa gataacatgg cttctttgcc agccacccac
      541 gagttgcaca tcttcggttc cattaacggt gtcgacttcg atatggtcgg tcaaggtacc
      601 ggtaacccaa acgacggtta cgaagaattg aacttgaagt ctaccaaggg cgacttgcag
      661 ttctcccctt ggattttggt tccacacatc ggttacggtt tccaccaata cttgccatac
      721 ccagacggta tgtccccatt ccaagccgcc atggttgacg gttctggtta ccaagtccac
      781 agaaccatgc aattcgaaga cggtgcttcc ttgaccgtga actacagata cacctacgaa
      841 ggttctcaca tcaagggcga agcccaagtt aagggtaccg gtttccctgc tgacggtcca
      901 gtcatgacta actccttgac cgctgctgac tggtgtcgtt ccaagaagac ctacccaaac
      961 gataagacca tcatttccac tttcaagtgg tcctacacca ccggtaacgg caagagatac
     1021 cgctctaccg ctcgtaccac ctacaccttc gccaagccaa tggctgccaa ctacttgaag
     1081 aaccagccta tgtacgtctt ccgtaagacc gagctgaagc attccaagac tgaattgaac
     1141 ttcaaggaat ggcaaaaggc tttcaccgac gtcatgggta tggacgaatt gtacaagtaa
     1201 gcggccgcct cgaggtcgac ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     1261 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1321 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     1381 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     1441 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt cacccggcca
     1501 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     1561 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     1621 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg
     1681 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     1741 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     1801 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg gtaaggaaaa
     1861 gactcacgtt tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1921 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1981 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2041 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2101 tatccgtact cctgatgatg catggttact caccactgcg atccccggca aaacagcatt
     2161 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2221 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2281 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2341 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc
     2401 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2461 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2521 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2581 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2641 cgatgagttt ttctaatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     2701 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     2761 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     2821 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     2881 cgccatccag tttaaacgag ctcgaattca tcgatgatat cagatccact agtggcctat
     2941 gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt
     3001 aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3061 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3421 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3661 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3841 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3901 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3961 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4021 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4081 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4141 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4201 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4261 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4321 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4381 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4441 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4501 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4561 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4621 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4681 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4741 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4801 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4861 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtct
//
