LOCUS       sequence-271062-        4428 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 4428)
  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 4428)
  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:hphMX4
FEATURES             Location/Qualifiers
     source          1..4428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..47)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            230..1805
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        230..573
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             574..1602
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             574..1602
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1608..1805
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1912..1931)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1913..1931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2018..2035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2189..2777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2269..2288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2948..3739,3740..3808))
                     /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     3571..3590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3809..3913)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3981..3999
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4037..4059)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4159..4178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4259..4277
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4259..4276
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      join(4396..4428,1..155)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
ORIGIN
        1 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
       61 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
      121 tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga taacagggta
      181 atatagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
      241 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
      301 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
      361 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcgg acctgcgagc agggaaacgc
      421 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
      481 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
      541 cagttctcac atcacatccg aacataaaca accatgggta aaaagcctga actcaccgcg
      601 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
      661 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
      721 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
      781 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
      841 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
      901 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
      961 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     1021 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     1081 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     1141 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     1201 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     1261 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     1321 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     1381 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     1441 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     1501 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     1561 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aatcagtact gacaataaaa
     1621 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     1681 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     1741 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     1801 tactgctgtc gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat
     1861 cgatgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt
     1921 gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg
     1981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3961 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4021 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4081 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4141 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca
     4201 tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat
     4261 ttaggtgaca ctatagaacg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga
     4321 tccccgggtt aattaagggg atagcttatc ctggttactt agactgttaa actaaggcgc
     4381 gccacttcta aataagcgaa tttcttatga tttatgattt ttattatt
//
