LOCUS       sequence-62215-e        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mWasabi with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4888)
  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 4888)
  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-yomWasabi-Kan
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(22..44)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     144..163
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        244..262
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     244..261
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1063..1250
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1118..1142)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1325..2681
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1325..1668
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1669..2478
                     /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(1736..1755)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2484..2681
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2785..2804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2786..2804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2891..2908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3062..3650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3142..3161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3821..4612,4613..4681))
                     /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     4444..4463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4682..4786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4854..4872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
       61 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      121 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      181 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
      241 acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg
      301 acggatcggt gacggtgctg gtttaattaa catggtaagt aaaggtgaag agacaactat
      361 gggggtaatc aaaccagaca tgaagatcaa actgaaaatg gaaggtaacg ttaatggcca
      421 cgcgtttgtt attgaaggtg aaggagaggg gaaaccttac gacggaacca acactatcaa
      481 tttggaggtt aaggaaggtg caccattacc attcagctat gacatcttaa caactgcatt
      541 ttcatacggc aacagagctt ttaccaagta ccctgacgat ataccaaatt acttcaaaca
      601 atcctttcct gaaggttatt catgggagag aacaatgacg tttgaagata agggcattgt
      661 caaggtcaaa tctgacattt ctatggaaga ggattcattc atctacgaga ttcacctcaa
      721 gggcgaaaac ttcccaccaa atggtccagt gatgcaaaag gaaactactg gatgggatgc
      781 ttctacagaa aggatgtacg tgagagatgg agtcttaaaa ggtgatgtta aaatgaagtt
      841 gctacttgaa ggtggtggcc atcatagagt tgatttcaaa acaatctaca gagccaaaaa
      901 ggctgtaaaa cttcctgatt atcattttgt ggatcatcgt atagaaattc taaaccacga
      961 caaagattac aataaggtca ctgtttatga aatagcagtt gccagaaatt ctacagatgg
     1021 aatggatgaa ttgtacaaat aaggcgcgcc acttctaaat aagcgaattt cttatgattt
     1081 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     1141 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     1201 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
     1261 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     1321 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     1381 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     1441 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     1501 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     1561 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     1621 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa
     1681 aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1741 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1801 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1861 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1921 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca
     1981 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     2041 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2101 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2161 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     2221 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2281 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2341 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2401 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2461 ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2521 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2581 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2641 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2701 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     2761 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2821 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2881 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3241 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3481 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4801 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4861 gtatcacgag gccctttcgt ctcgcgcg
//
