LOCUS       sequence-271023-        4117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal A1-FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4117)
  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 4117)
  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-A1-FLAG:HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             229..927
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      931..1128
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1232..1251)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1233..1251)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1338..1355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1509..2097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1589..1608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2268..3059,3060..3128))
                     /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     2891..2910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3129..3233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3301..3319
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3357..3379)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3479..3498
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3579..3597
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3579..3596
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3693..3716
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3740..3927
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3795..3819)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        join(4002..4117,1..228)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
       61 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
      121 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
      181 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat ggcagaacca
      241 gcccaaaaaa agcaaaaaca aactgttcag gagcgcaagg cgtttatctc ccgtatcact
      301 aatgaaacta aaattcaaat cgctatttcg ctgaatggtg gttatattca aataaaagat
      361 tcgattcttc ctgcaaagaa ggatgacgat gtagcttccc aagctactca gtcacaggtc
      421 atcgatattc acacaggtgt tggctttttg gatcatatga tccatgcgtt ggcaaaacac
      481 tctggttggt ctcttattgt tgaatgtatt ggtgacctgc acattgacga tcaccatact
      541 accgaagatt gcggtatcgc attagggcaa gcgttcaaag aagcaatggg tgctgtccgt
      601 ggtgtaaaaa gattcggtac tgggttcgca ccattggatg aggcgctatc acgtgccgta
      661 gtcgatttat ctagtagacc atttgctgta atcgaccttg gattgaagag agagatgatt
      721 ggtgatttat ccactgaaat gattccacac tttttggaaa gtttcgcgga ggcggccaga
      781 attactttgc atgttgattg tctgagaggt ttcaacgatc accacagaag tgagagtgcg
      841 ttcaaggctt tggctgttgc cataagagaa gctatttcta gcaatggcac caatgacgtt
      901 ccctcaacca aaggtgtttt gatgtgaagt actgacaata aaaagattct tgttttcaag
      961 aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc
     1021 gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag
     1081 aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga
     1141 tactaacgcc gccatccagt ttaaacgagc tcgaattcat cgatgatatc agatccacta
     1201 gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata
     1261 agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1321 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1381 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1441 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1501 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1561 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1621 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1681 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1741 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1801 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1861 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1921 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1981 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2041 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2101 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2161 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2221 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2281 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2341 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2401 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2461 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2521 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2581 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2641 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2701 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2761 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2821 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2881 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2941 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3001 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3061 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3121 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3181 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3241 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3301 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3361 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3421 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     3481 gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc
     3541 tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaacg
     3601 cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt aattaagggg
     3661 atagcctgga catgctcgag tggtctctga tggattacaa ggacgacgat gacaagtaag
     3721 gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt
     3781 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     3841 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt
     3901 cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg taatatagat
     3961 ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata
     4021 ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca
     4081 tttagcccat acatccccat gtataatcat ttgcatc
//
