LOCUS       sequence-62147-e        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized EGFP
            with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  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 4894)
  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-Kan
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..85
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        166..184
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     166..183
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(254..256,257..967)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     terminator      991..1178
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1046..1070)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1253..2609
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1253..1596
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1597..2406
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1664..1683)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2412..2609
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2713..2732)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2714..2732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2819..2836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2990..3578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3070..3089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3749..4540,4541..4609))
                     /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     4372..4391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4610..4714)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4782..4800
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4838..4860)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
       61 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga
      121 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
      181 tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcg gtgacggtgc
      241 tggtttaatt aacatgtcaa aaggcgagga actattcaca ggtgtcgtac caatactggt
      301 agaattggat ggcgacgtaa acgggcataa gttttctgtg agtggtgagg gtgaagggga
      361 cgcaacttat ggcaaactta cacttaagtt catctgcacc acagggaagc tgcctgtccc
      421 atggccaact ctagttacaa ctctcactta cggagtacaa tgtttttctc gatacccaga
      481 ccacatgaaa caacatgact ttttcaaatc tgccatgcct gagggatatg tgcaagagag
      541 aactattttc tttaaagatg atggtaatta caaaactcgt gctgaagtga agtttgaagg
      601 agatacactt gttaatagaa ttgagttaaa gggtattgac tttaaggaag atggtaatat
      661 cttaggtcac aaattggaat acaactacaa ttcacataac gtttacatca tggcagataa
      721 acagaaaaac ggcataaaag ttaatttcaa aattagacat aatatcgaag atggttccgt
      781 tcaactcgct gatcattatc aacaaaatac accaatcgga gatggacctg ttctattgcc
      841 agacaaccat tacttatcta cacagagcgc cttgtcaaaa gatcctaatg aaaagagaga
      901 tcacatggtt ttgttagaat ttgtcaccgc tgcgggtata acgttgggca tggatgaatt
      961 atacaagtaa ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt
     1021 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
     1081 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
     1141 tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg
     1201 gtaatataga tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga
     1261 ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg
     1321 tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca tacattttga
     1381 tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa
     1441 cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa
     1501 aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg
     1561 atacagttct cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt
     1621 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1681 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     1741 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1801 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1861 cctgatgatg catggttact caccactgcg atccccggca aaacagcatt ccaggtatta
     1921 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1981 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2041 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2101 aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg
     2161 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2221 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2281 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2341 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2401 ttctaatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt
     2461 ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc
     2521 gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca
     2581 atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag
     2641 tttaaacgag ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg
     2701 atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt
     2761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3181 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3421 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3541 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3601 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3661 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3721 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3781 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3841 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3901 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3961 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4021 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4081 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4141 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4201 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4261 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4321 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4381 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4441 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4501 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4561 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4621 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4681 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4741 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4801 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4861 agacggtcac agcttgtctg taagcggatg ccgg
//
