LOCUS       40924_48198        4905 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid with a hygromycin resistance 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 4905)
  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 4905)
  AUTHORS   EUROSCARF
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4905)
  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..4905
                     /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"
     promoter        853..1196
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1203..2222
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     terminator      2279..2468
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(2543..2561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3581..4438)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4439..4543)
                     /label=AmpR promoter
     promoter        4889..4905
                     /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 gcgcgccaga tctgtttagc ttgcctcgtc cccgccgggt
      841 cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc
      901 tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca
      961 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
     1021 cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
     1081 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
     1141 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg
     1201 gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     1261 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     1321 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     1381 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     1441 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     1501 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     1561 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     1621 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     1681 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     1741 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     1801 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     1861 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     1921 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     1981 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     2041 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     2101 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     2161 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     2221 aataatctcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     2281 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     2341 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     2401 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     2461 gctcgaaggc tttaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     2521 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     2581 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4561 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4621 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4681 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4741 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4801 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     4861 acataacctt atgtatcata cacatacgat ttaggtgaca ctata
//
