LOCUS       sequence-271037-        3996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal S6-FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3996)
  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 3996)
  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-FLAG:natMX4
FEATURES             Location/Qualifiers
     source          1..3996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(19..123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     191..209
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(247..269)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     369..388
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        469..487
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     469..486
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             583..606
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      630..817
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(685..709)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            892..2011
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        892..1235
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1236..1808
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1814..2011
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2118..2137)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2119..2137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2224..2241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2395..2983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2475..2494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3154..3945,3946..3996,1..18))
                     /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     3777..3796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
       61 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      121 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      181 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
      241 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
      301 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      361 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
      421 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
      481 ctatagaacg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt
      541 aattaagggg atagcttatc ctggttactt agactgttaa acgattacaa ggacgacgat
      601 gacaagtaag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
      661 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
      721 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
      781 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
      841 taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag
      901 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
      961 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     1021 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac
     1081 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
     1141 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
     1201 tacagttctc acatcacatc cgaacataaa caaccatggg taccactctt gacgacacgg
     1261 cttaccggta ccgcaccagt gtcccggggg acgccgaggc catcgaggca ctggatgggt
     1321 ccttcaccac cgacaccgtc ttccgcgtca ccgccaccgg ggacggcttc accctgcggg
     1381 aggtgccggt ggacccgccc ctgaccaagg tgttccccga cgacgaatcg gacgacgaat
     1441 cggacgacgg ggaggacggc gacccggact cccggacgtt cgtcgcgtac ggggacgacg
     1501 gcgacctggc gggcttcgtg gtcgtctcgt actccggctg gaaccgccgg ctgaccgtcg
     1561 aggacatcga ggtcgccccg gagcaccggg ggcacggggt cgggcgcgcg ttgatggggc
     1621 tcgcgacgga gttcgcccgc gagcggggcg ccgggcacct ctggctggag gtcaccaacg
     1681 tcaacgcacc ggcgatccac gcgtaccggc ggatggggtt caccctctgc ggcctggaca
     1741 ccgccctgta cgacggcacc gcctcggacg gcgagcaggc gctctacatg agcatgccct
     1801 gcccctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1861 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1921 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     1981 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     2041 agtgtcgaaa acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc
     2101 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     2161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3961 gcaaaatgcc gcaaaaaagg gaataagggc gacacg
//
