LOCUS       sequence-62213-e        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            LSS-mKate2 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4876)
  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 4876)
  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-yoLSS-mKate2-Kan
FEATURES             Location/Qualifiers
     source          1..4876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        215..233
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     215..232
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1022..1209
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1077..1101)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1284..2640
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1284..1627
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1628..2437
                     /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(1695..1714)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2443..2640
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2744..2763)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2745..2763)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2850..2867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3021..3609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3101..3120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3780..4571,4572..4640))
                     /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     4403..4422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4641..4745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4813..4831
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4869..4876,1..15))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
       61 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      121 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
      181 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaacgcggc
      241 cgccagctga agcttcgtac gctgcaggtc gacggatcgg tgacggtgct ggtttaatta
      301 acatggtttc agaattgatc aaggaaaata tgcacatgaa actgtatatg gaaggtacag
      361 ttaacaacca tcattttaag tgtacctcag aaggagaagg gaaaccatac gaaggcactc
      421 agactatgag aattaaagtg gtcgaaggtg gtccactacc atttgctttt gacatcctcg
      481 ctacatcttt catgtacggt tcctacactt tcattaatca cacacaaggc ataccagact
      541 ttttcaaaca atcatttcct gagggtttta cttgggaaag agtaaccact tatgaagatg
      601 ggggtgtact gacagctaca caagatacat ctttacaaga tggatgtcta atctacaatg
      661 ttaagattag aggagttaac ttcacatcta atggtcctgt catgcaaaag aaaacgttag
      721 gctgggaggc aggtacagaa atgctttatc cagcagacgg cggtttggaa ggtagaagtg
      781 acgatgcttt gaaattggta gggggaggcc atcttatatg caacctaaaa tctacttaca
      841 gaagcaaaaa gcctgccaag aatttgaaag ttccaggcgt ctactatgtt gatcgtaggt
      901 tagagcgaat caaggaggcg gataaggaaa cctacgtgga acagcatgag gtggcagtag
      961 ccagatactg tgatcttcct tccaaattag gacacagata aggcgcgcca cttctaaata
     1021 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
     1081 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
     1141 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
     1201 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
     1261 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     1321 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     1381 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     1441 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     1501 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     1561 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     1621 aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga
     1681 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1741 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1801 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1861 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1921 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1981 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2041 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2101 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2161 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2221 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2281 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2341 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2401 attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt
     2461 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     2521 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     2581 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     2641 ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc atcgatgata
     2701 tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta
     2761 ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc
     2821 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2881 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3181 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     3241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3421 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3661 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3721 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3781 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3841 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3901 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3961 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4021 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4081 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4141 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4201 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4261 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4321 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4381 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4441 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4501 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4561 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4621 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4681 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4741 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4801 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4861 ggtgaaaacc tctgac
//
