LOCUS       sequence-271058-        3972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal S6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3972)
  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 3972)
  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-S6:natMX4
FEATURES             Location/Qualifiers
     source          1..3972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..34
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(72..94)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     194..213
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        294..312
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     294..311
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      431..618
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(486..510)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            693..1812
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        693..1036
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1037..1609
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1615..1812
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1919..1938)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1920..1938)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2025..2042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2196..2784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2276..2295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2955..3746,3747..3815))
                     /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     3578..3597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3816..3920)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
       61 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
      121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
      181 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg
      241 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg
      301 tgacactata gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc
      361 gggttaatta aggggatagc ttatcctggt tacttagact gttaaactaa ggcgcgccac
      421 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
      481 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
      541 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
      601 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc
      661 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
      721 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
      781 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
      841 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
      901 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
      961 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     1021 ccgaacataa acaaccatgg gtaccactct tgacgacacg gcttaccggt accgcaccag
     1081 tgtcccgggg gacgccgagg ccatcgaggc actggatggg tccttcacca ccgacaccgt
     1141 cttccgcgtc accgccaccg gggacggctt caccctgcgg gaggtgccgg tggacccgcc
     1201 cctgaccaag gtgttccccg acgacgaatc ggacgacgaa tcggacgacg gggaggacgg
     1261 cgacccggac tcccggacgt tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt
     1321 ggtcgtctcg tactccggct ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc
     1381 ggagcaccgg gggcacgggg tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg
     1441 cgagcggggc gccgggcacc tctggctgga ggtcaccaac gtcaacgcac cggcgatcca
     1501 cgcgtaccgg cggatggggt tcaccctctg cggcctggac accgccctgt acgacggcac
     1561 cgcctcggac ggcgagcagg cgctctacat gagcatgccc tgcccctaat cagtactgac
     1621 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     1681 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     1741 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     1801 gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa aacgagctcg
     1861 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     1921 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     1981 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2701 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2761 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2821 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2881 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2941 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3001 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3061 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3121 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3181 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3241 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3301 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3361 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3421 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3481 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3541 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3601 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3661 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3721 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3781 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3841 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3901 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     3961 attattatca tg
//
