LOCUS       sequence-159997-        4726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Useful for tagging yeast proteins with C-terminal YFP tag; his 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   Hailey DW, Davis TN, Muller EG
  TITLE     Fluorescence resonance energy transfer using color variants of green
            fluorescent protein.
  JOURNAL   Methods Enzymol. 2002;351:34-49.
  PUBMED    12073355
REFERENCE   2  (bases 1 to 4726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Enzymol."; date: "2002"; volume: "351"; pages: "34-49"
COMMENT     Sequence Label: pDH5
FEATURES             Location/Qualifiers
     source          1..4726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..776
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(90..118)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     713..733
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      797..984
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(852..876)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1059..2259
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1059..1402
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1403..2056
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2062..2259
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2363..2382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2364..2382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2469..2486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2640..3228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2720..2739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3399..4190,4191..4259))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4022..4041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4260..4364)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4432..4450
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4488..4510)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4610..4629
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4710..4726,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4710..4726,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      121 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
      181 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
      241 tcactacttt cggttatggt ctaaaatgct ttgctagata cccagatcat atgaaacggc
      301 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
      361 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
      421 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
      481 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
      541 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc
      601 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
      661 tgtcctatca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
      721 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaatagggcg
      781 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat
      841 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
      901 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc
      961 tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg
     1021 tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc
     1081 tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt
     1141 agcccataca tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc
     1201 gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag
     1261 acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc
     1321 cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca
     1381 tcacatccga acataaacaa ccatgggtag gagggctttt gtagaaagaa atacgaacga
     1441 aacgaaaatc agcgttgcca tcgctttgga caaagctccc ttacctgaag agtcgaattt
     1501 tattgatgaa cttataactt ccaagcatgc aaaccaaaag ggagaacaag taatccaagt
     1561 agacacggga attggattct tggatcacat gtatcatgca ctggctaaac atgcaggctg
     1621 gagcttacga ctttactcaa gaggtgattt aatcatcgat gatcatcaca ctgcagaaga
     1681 tactgctatt gcacttggta ttgcattcaa gcaggctatg ggtaactttg ccggcgttaa
     1741 aagatttgga catgcttatt gtccacttga cgaagctctt tctagaagcg tagttgactt
     1801 gtcgggacgg ccctatgctg ttatcgattt gggattaaag cgtgaaaagg ttggggaatt
     1861 gtcctgtgaa atgatccctc acttactata ttccttttcg gtagcagctg gaattacttt
     1921 gcatgttacc tgcttatatg gtagtaatga ccatcatcgt gctgaaagcg cttttaaatc
     1981 tctggctgtt gccatgcgcg cggctactag tcttactgga agttctgaag tcccaagcac
     2041 gaagggagtg ttgtaaagag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     2101 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     2161 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     2221 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     2281 cgccatccag tttaaacgag ctcgaattca tcgatgatat cagatccact agtggcctat
     2341 gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt
     2401 aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2461 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2521 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2581 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2641 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2701 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2761 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2821 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2881 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2941 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3001 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3061 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3121 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3181 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3241 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3301 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3361 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3421 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3481 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3541 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3601 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3661 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3721 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3781 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3841 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3901 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3961 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4021 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4081 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4141 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4201 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4261 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4321 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4381 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4441 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4501 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4561 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4621 cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa
     4681 tacataacct tatgtatcat acacatacga tttaggtgac actata
//
