LOCUS       sequence-62225-e        4864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            Clover with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4864)
  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 4864)
  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-yoClover-Kan
FEATURES             Location/Qualifiers
     source          1..4864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..25
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(63..85)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     185..204
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        285..303
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     285..302
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1080..1267
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1135..1159)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1342..2698
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1342..1685
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1686..2495
                     /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(1753..1772)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2501..2698
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2802..2821)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2803..2821)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2908..2925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3079..3667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3159..3178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3838..4629,4630..4698))
                     /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     4461..4480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4699..4803)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
       61 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
      121 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
      181 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
      241 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
      301 agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacggatcgg tgacggtgct
      361 ggtttaatta acatggtttc taaaggtgaa gagctattca ctggagtagt tccaattctg
      421 gttgaactag atggcgacgt gaatggccac aaattttccg tgcgtgggga aggcgaaggt
      481 gatgcaacta acggtaaact gacattaaag tttatctgta ccactggtaa gttgcctgta
      541 ccatggccaa cgttagttac aactttcggg tatggtgtag cttgcttttc aaggtaccca
      601 gaccacatga aacaacatga tttctttaag tctgcaatgc ctgaaggtta cgttcaggag
      661 agaacaatct cttttaaaga cgatggcact tacaaaacaa gagctgaagt aaagtttgaa
      721 ggagatacac ttgtcaatag aatagaattg aaaggtattg atttcaaaga ggatggaaat
      781 attctaggcc ataagttaga atacaacttc aactcacaca atgtttacat tactgcagat
      841 aaacaaaaga atggtataaa agccaacttt aagatcagac ataacgtgga ggatggttct
      901 gtccaactcg ctgaccatta ccaacaaaat accccaatcg gagatggacc agtccttttg
      961 cctgacaatc attatttgag tcatcagagc gctttatcaa aagaccctaa tgaaaagaga
     1021 gatcacatgg ttttgctcga atttgtcaca gccgcgtaag gcgcgccact tctaaataag
     1081 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     1141 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     1201 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     1261 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
     1321 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     1381 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     1441 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     1501 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     1561 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     1621 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     1681 caaccatggg taaggaaaag actcacgttt cgaggccgcg attaaattcc aacatggatg
     1741 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1801 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1861 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1921 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1981 tccccggcaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2041 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2101 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2161 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2221 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2281 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2341 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2401 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2461 tgcagtttca tttgatgctc gatgagtttt tctaatcagt actgacaata aaaagattct
     2521 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     2581 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     2641 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     2701 gtcgattcga tactaacgcc gccatccagt ttaaacgagc tcgaattcat cgatgatatc
     2761 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     2821 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4801 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4861 aacc
//
