LOCUS       sequence-62145-e        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mTagBFP with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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-yomTagBFP-Kan
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     232..251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(470..574)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     642..660
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(698..720)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     820..839
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        920..938
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     920..937
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1724..1911
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1779..1803)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1986..3342
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1986..2329
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2330..3139
                     /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(2397..2416)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3145..3342
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3446..3465)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3447..3465)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3552..3569)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3723..4311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3803..3822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4482..4873,1..400,401..469))
                     /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"
ORIGIN
        1 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
       61 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
      181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
      241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
      301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
      361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
      481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
      601 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      661 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      721 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      781 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      841 tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct
      901 tatgtatcat acacatacga tttaggtgac actatagaac gcggccgcca gctgaagctt
      961 cgtacgctgc aggtcgacgg atcggtgacg gtgctggttt aattaacatg agtgagttga
     1021 ttaaggaaaa catgcacatg aagttgtaca tggaaggaac agtagataat catcatttca
     1081 aatgtacgtc agaaggtgag ggcaaacctt atgaaggaac tcaaactatg agaattaaag
     1141 tcgttgaggg tggcccacta ccatttgcat tcgatatcct cgcaacatct tttctgtatg
     1201 gttctaaaac ctttatcaac catacacaag gcataccaga tttctttaag caatcatttc
     1261 ctgaaggttt cacatgggaa agagtaacta cctatgaaga tggtggtgtc cttacagcta
     1321 ctcaagacac gtctttgcag gatggttgtt taatctacaa tgttaagata agaggagtga
     1381 actttacatc caatggacca gtcatgcaaa agaaaactct aggttgggag gcattcacag
     1441 aaactttgta cccagccgat gggggcttag aagggagaaa tgacatggct ttaaaactgg
     1501 ttggcggtag ccatttgatt gcgaacatca aaacaaccta cagaagcaaa aagccagcta
     1561 aaaatcttaa gatgcctgta tactacgtgg attacagatt agaaaggaag gaagctaata
     1621 atgaaactta cgttgaacag cacgaggtgg ccgttgcccg ttattgcgac cttccttcaa
     1681 aattgggaca caaactaaac taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     1741 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     1801 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     1861 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     1921 tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     1981 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     2041 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     2101 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     2161 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     2221 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     2281 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgggtaagga
     2341 aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2401 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     2461 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2521 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2581 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gcaaaacagc
     2641 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2701 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2761 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2821 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
     2881 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2941 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3001 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3061 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3121 gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     3181 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     3241 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     3301 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     3361 cgccgccatc cagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     3421 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     3481 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3541 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3901 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4141 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4321 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4381 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4441 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4501 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4561 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4621 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4681 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4741 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4801 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4861 tcaaggcgag tta
//
