LOCUS       Exported                4899 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 4899)
  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 4899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4899
                     /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..1195
                     /locus_tag="ymBaojin"
                     /label=ymBaojin
     terminator      1233..1420
                     /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(1288..1312)
                     /locus_tag="pHybLex-R"
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1495..2851
                     /locus_tag="kanMX"
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        1495..1838
                     /locus_tag="TEF promoter"
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1839..2648
                     /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(1906..1925)
                     /locus_tag="Kan-R"
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2654..2851
                     /locus_tag="TEF terminator"
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2955..2974)
                     /locus_tag="T7"
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2956..2974)
                     /locus_tag="T7 promoter"
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3061..3078)
                     /locus_tag="L4440"
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3232..3820)
                     /direction=LEFT
                     /locus_tag="ori"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3312..3331)
                     /locus_tag="pBR322ori-F"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3991..4851)
                     /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     4614..4633
                     /locus_tag="Amp-R"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4852..4899,1..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 atggtttcta agggtgaaga agaaaacatg gcttctactc cattcaagtt
      541 ccaattgaag ggtactatta acggtaagtc tttcactgtt gaaggtgaag gtgaaggtaa
      601 ctctcatgaa ggttctcaca agggtaagta cgtttgtact tctggtaagt tgccaatgtc
      661 ttgggctgct ttgggtacta ctttcggtta cggtatgaag tactacacta agtacccatc
      721 tggtttaaag aactggttca gagaagttat gccaggtggt ttcacttacg atagacatat
      781 tcaatacaag ggtgatggtt ctattcatgc taagcatcaa catttcatga agaacggtac
      841 ttaccataac attgttgaat tcactggtca agatttcaag gaaaattctc cagttttgac
      901 cggtgatatg aacgtttctt tgccaaatga agttccacaa attccaagag atgatggtgt
      961 tgaatgtcca gttactttat tgtacccatt gttgtctgat aagtctaagt atgttgaagc
     1021 tcatcaatac actatttgta agccattgca taaccaacca gctccagatg tcccatacca
     1081 ttggattaga aagcaataca ctcaatctaa ggatgatgct gaagaaagag atcatatttg
     1141 tcaatctgaa actttagaag ctcatttgaa gggtatggat gaattgtaca agtaagcggc
     1201 cgcctcgagg tcggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     1261 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     1321 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     1381 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     1441 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
     1501 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
     1561 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
     1621 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
     1681 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     1741 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     1801 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
     1861 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1921 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     1981 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     2041 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2101 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
     2161 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     2221 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     2281 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     2341 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     2401 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2461 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2521 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2581 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2641 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     2701 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     2761 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     2821 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     2881 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     2941 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     3001 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3061 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3421 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3661 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3961 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4021 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4081 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4141 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4201 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4261 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4321 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4381 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4441 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4501 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4561 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4621 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4681 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4741 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4801 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4861 ttttcaatat tattgaagca tttatcaggg ttattgtct
//
