LOCUS       sequence-62216-e        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            Emerald 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-yoEmerald-Kan
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        124..142
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     124..141
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      949..1136
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1004..1028)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1211..2567
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1211..1554
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1555..2364
                     /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(1622..1641)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2370..2567
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2671..2690)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2672..2690)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2777..2794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2948..3536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3028..3047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3707..4498,4499..4567))
                     /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     4330..4349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4568..4672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4740..4758
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4796..4818)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
       61 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
      121 acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg
      181 acggatcggt gacggtgctg gtttaattaa catgtcaaaa ggcgaggaac tattcacagg
      241 tgtcgtacca atactggtag aattggatgg cgacgtaaac gggcataagt tttctgtgag
      301 tggtgagggt gaaggggacg caacttatgg caaacttaca cttaagttca tctgcaccac
      361 agggaagctg cctgtcccat ggccaactct agttacaact ctcacttacg gagtacaatg
      421 ttttgcacga tacccagacc acatgaaaca acatgacttt ttcaaatctg ccatgcctga
      481 gggatatgtg caagagagaa ctattttctt taaagatgat ggtaattaca aaactcgtgc
      541 tgaagtgaag tttgaaggag atacacttgt taatagaatt gagttaaagg gtattgactt
      601 taaggaagat ggtaatatct taggtcacaa attggaatac aactacaatt cacataaggt
      661 ttacatcaca gcagataaac agaaaaacgg cataaaagtt aatttcaaaa ccagacataa
      721 tatcgaagat ggttccgttc aactcgctga tcattatcaa caaaatacac caatcggaga
      781 tggacctgtt ctattgccag acaaccatta cttatctaca cagagcgcct tgtcaaaaga
      841 tcctaatgaa aagagagatc acatggtttt gttagaattt gtcaccgctg cgggtataac
      901 gttgggcatg gatgaattat acaagtaagg cgcgccactt ctaaataagc gaatttctta
      961 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     1021 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     1081 aggttgcttt ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga
     1141 tccgctaggg ataacagggt aatatagatc tgtttagctt gcctcgtccc cgccgggtca
     1201 cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc
     1261 aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt
     1321 tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca
     1381 gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc
     1441 cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc
     1501 ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt
     1561 aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1621 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     1681 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1741 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1801 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
     1861 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1921 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1981 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2041 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag
     2101 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2161 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2221 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2281 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2341 ttgatgctcg atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga
     2401 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     2461 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     2521 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     2581 actaacgccg ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag
     2641 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     2701 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3121 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     3181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3361 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4681 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4741 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4801 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4861 aagcccgtca gggcgcgtca gcgggtgttg gcgg
//
