LOCUS       sequence-62007-e        4738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized EGFP
            with SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4738)
  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 4738)
  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: "PLoS One.";
            date: "2013-07-2"; volume: "8(7):e67902. doi"; pages: " 
            10.1371/journal.pone.0067902. Print 2013"
COMMENT     Sequence Label: pFA6a-link-yoEGFP-SpHis5
FEATURES             Location/Qualifiers
     source          1..4738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..45
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(83..105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     205..224
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        305..323
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     305..322
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(393..395,396..1106)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     terminator      1130..1317
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1185..1209)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1392..2592
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1392..1735
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1736..2389
                     /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      2395..2592
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2696..2715)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2697..2715)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2802..2819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2973..3561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3053..3072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3732..4523,4524..4592))
                     /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     4355..4374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4593..4697)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
       61 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
      121 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      181 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      241 tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca
      301 tacgatttag gtgacactat agaacgcggc cgccagctga agcttcgtac gctgcaggtc
      361 gacggatcgg tgacggtgct ggtttaatta acatgtcaaa aggcgaggaa ctattcacag
      421 gtgtcgtacc aatactggta gaattggatg gcgacgtaaa cgggcataag ttttctgtga
      481 gtggtgaggg tgaaggggac gcaacttatg gcaaacttac acttaagttc atctgcacca
      541 cagggaagct gcctgtccca tggccaactc tagttacaac tctcacttac ggagtacaat
      601 gtttttctcg atacccagac cacatgaaac aacatgactt tttcaaatct gccatgcctg
      661 agggatatgt gcaagagaga actattttct ttaaagatga tggtaattac aaaactcgtg
      721 ctgaagtgaa gtttgaagga gatacacttg ttaatagaat tgagttaaag ggtattgact
      781 ttaaggaaga tggtaatatc ttaggtcaca aattggaata caactacaat tcacataacg
      841 tttacatcat ggcagataaa cagaaaaacg gcataaaagt taatttcaaa attagacata
      901 atatcgaaga tggttccgtt caactcgctg atcattatca acaaaataca ccaatcggag
      961 atggacctgt tctattgcca gacaaccatt acttatctac acagagcgcc ttgtcaaaag
     1021 atcctaatga aaagagagat cacatggttt tgttagaatt tgtcaccgct gcgggtataa
     1081 cgttgggcat ggatgaatta tacaagtaag gcgcgccact tctaaataag cgaatttctt
     1141 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     1201 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     1261 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     1321 atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc
     1381 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     1441 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     1501 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     1561 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
     1621 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
     1681 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
     1741 taggagggct tttgtagaaa gaaatacgaa cgaaacgaaa atcagcgttg ccatcgcttt
     1801 ggacaaagct cccttacctg aagagtcgaa ttttattgat gaacttataa cttccaagca
     1861 tgcaaaccaa aagggagaac aagtaatcca agtagacacg ggaattggat tcttggatca
     1921 catgtatcat gcactggcta aacatgcagg ctggagctta cgactttact caagaggtga
     1981 tttaatcatc gatgatcatc acactgcaga agatactgct attgcacttg gtattgcatt
     2041 caagcaggct atgggtaact ttgccggcgt taaaagattt ggacatgctt attgtccact
     2101 tgacgaagct ctttctagaa gcgtagttga cttgtcggga cggccctatg ctgttatcga
     2161 tttgggatta aagcgtgaaa aggttgggga attgtcctgt gaaatgatcc ctcacttact
     2221 atattccttt tcggtagcag ctggaattac tttgcatgtt acctgcttat atggtagtaa
     2281 tgaccatcat cgtgctgaaa gcgcttttaa atctctggct gttgccatgc gcgcggctac
     2341 tagtcttact ggaagttctg aagtcccaag cacgaaggga gtgttgtaaa gagtactgac
     2401 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     2461 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     2521 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     2581 gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat
     2641 tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta
     2701 tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc
     2761 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2821 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2881 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2941 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3001 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3061 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     3181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3661 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3721 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3781 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3841 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3901 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3961 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4021 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4081 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4141 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4201 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4261 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4321 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4381 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4441 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4501 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4561 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4621 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4681 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaacca
//
