LOCUS       sequence-258437-        3830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal SRS-FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3830)
  AUTHORS   Nemec AA, Tomko RJ Jr
  TITLE     A suite of PCR-based peptide tagging plasmids to permit 
            epitope-targeted enzymatic functionalization of yeast proteins.
  JOURNAL   Yeast. 2020 May 13. doi: 10.1002/yea.3471.
  PUBMED    32401365
REFERENCE   2  (bases 1 to 3830)
  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: "Yeast. 2020
            May 13. doi: 10.1002/yea.3471."
COMMENT     Sequence Label: pFA6a-SRS-FLAG:HIS3MX6
FEATURES             Location/Qualifiers
     source          1..3830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             83..106
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     gene            163..1363
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        163..506
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             507..1160
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      1166..1363
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1467..1486)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1468..1486)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1573..1590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1744..2332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1824..1843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2503..3294,3295..3363))
                     /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     3126..3145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3364..3468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3536..3554
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3592..3614)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3714..3733
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3814..3830,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3814..3830,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 actgcccgaa accggtggtg gcgattacaa ggacgacgat gacaagtaag gcgcgccaga
      121 tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
      181 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
      241 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
      301 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
      361 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
      421 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
      481 cacatcacat ccgaacataa acaaccatgg gtaggagggc ttttgtagaa agaaatacga
      541 acgaaacgaa aatcagcgtt gccatcgctt tggacaaagc tcccttacct gaagagtcga
      601 attttattga tgaacttata acttccaagc atgcaaacca aaagggagaa caagtaatcc
      661 aagtagacac gggaattgga ttcttggatc acatgtatca tgcactggct aaacatgcag
      721 gctggagctt acgactttac tcaagaggtg atttaatcat cgatgatcat cacactgcag
      781 aagatactgc tattgcactt ggtattgcat tcaagcaggc tatgggtaac tttgccggcg
      841 ttaaaagatt tggacatgct tattgtccac ttgacgaagc tctttctaga agcgtagttg
      901 acttgtcggg acggccctat gctgttatcg atttgggatt aaagcgtgaa aaggttgggg
      961 aattgtcctg tgaaatgatc cctcacttac tatattcctt ttcggtagca gctggaatta
     1021 ctttgcatgt tacctgctta tatggtagta atgaccatca tcgtgctgaa agcgctttta
     1081 aatctctggc tgttgccatg cgcgcggcta ctagtcttac tggaagttct gaagtcccaa
     1141 gcacgaaggg agtgttgtaa agagtactga caataaaaag attcttgttt tcaagaactt
     1201 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     1261 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     1321 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
     1381 acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
     1441 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
     1501 ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1561 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1681 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1921 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2161 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3481 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3541 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3601 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3661 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3721 agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa
     3781 ttaatacata accttatgta tcatacacat acgatttagg tgacactata
//
