LOCUS       40924_48208        4749 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid with a HIS3MX6 marker that provides a template for PCR to 
            fuse yeast enhanced GFP (yeGFP) to the C-terminus of a protein of 
            interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4749)
  AUTHORS   Janke C, Magiera MM, Rathfelder N, Taxis C, Reber S, Maekawa H, 
            Moreno-Borchart A, Doenges G, Schwob E, Schiebel E, Knop M.
  TITLE     A versatile toolbox for PCR-based tagging of yeast genes: new 
            fluorescent proteins, more markers and promoter substitution 
            cassettes.
  JOURNAL   Yeast 2004;21:947-62.
  PUBMED    15334558
REFERENCE   2  (bases 1 to 4749)
  AUTHORS   EUROSCARF
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4749)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2004"; volume: "21"; pages: "947-62"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..795
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="LSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKTDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      820..1007
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     gene            1082..2282
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        complement(2387..2405)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2663..3251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3425..4282)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4283..4387)
                     /label=AmpR promoter
     promoter        4733..4749
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatccgg agcaggtgct
       61 ggtgctggtg ctggagcaat tctgtctaaa ggtgaagaat tattcactgg tgttgtccca
      121 attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt
      181 gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg
      241 ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg ttttgcgaga
      301 tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt
      361 caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag
      421 tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat
      481 ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg
      541 gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat
      601 ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc
      661 ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa
      721 aagacagacc acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg
      781 gatgaattgt acaaataact gcgcgccact tctaaataag cgaatttctt atgatttatg
      841 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
      901 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
      961 tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg
     1021 gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag
     1081 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     1141 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat
     1201 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga
     1261 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     1321 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     1381 cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg taggagggct
     1441 tttgtagaaa gaaatacgaa cgaaacgaaa atcagcgttg ccatcgcttt ggacaaagct
     1501 cccttacctg aagagtcgaa ttttattgat gaacttataa cttccaagca tgcaaaccaa
     1561 aagggagaac aagtaatcca agtagacacg ggaattggat tcttggatca catgtatcat
     1621 gcactggcta aacatgcagg ctggagctta cgactttact caagaggtga tttaatcatc
     1681 gatgatcatc acactgcaga agatactgct attgcacttg gtattgcatt caagcaggct
     1741 atgggtaact ttgccggcgt taaaagattt ggacatgctt attgtccact tgacgaagct
     1801 ctttctagaa gcgtagttga cttgtcggga cggccctatg ctgttatcga tttgggatta
     1861 aagcgtgaaa aggttgggga attgtcctgt gaaatgatcc ctcacttact atattccttt
     1921 tcggtagcag ctggaattac tttgcatgtt acctgcttat atggtagtaa tgaccatcat
     1981 cgtgctgaaa gcgcttttaa atctctggct gttgccatgc gcgcggctac tagtcttact
     2041 ggaagttctg aagtcccaag cacgaaggga gtgttgtaaa gagtactgac aataaaaaga
     2101 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2161 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2221 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     2281 tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga
     2341 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     2401 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     2461 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2521 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2581 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2641 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2701 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2761 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2821 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     2881 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2941 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3001 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3061 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3121 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3181 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3241 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3301 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3361 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3421 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3481 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3541 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3601 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3661 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3721 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3781 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3841 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3901 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3961 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4021 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4081 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4141 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4201 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4261 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4321 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4381 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4441 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     4501 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4561 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     4621 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
     4681 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cgatttaggt
     4741 gacactata
//
