LOCUS       sequence-271011-        4206 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 4206)
  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 4206)
  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:kanMX6
FEATURES             Location/Qualifiers
     source          1..4206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..24)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(684..1475,1476..1544))
                     /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     1307..1326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1545..1649)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1717..1735
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1773..1795)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1895..1914
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1995..2013
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1995..2012
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      2132..2319
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2187..2211)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2394..3750
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2394..2737
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2738..3547
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2805..2824)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3553..3750
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3854..3873)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3855..3873)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3960..3977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4131..4206,1..513))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
       61 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      121 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      181 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      241 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      301 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      361 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      421 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      481 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      541 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      601 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      661 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      721 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      781 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      841 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      901 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      961 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1021 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1081 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1141 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1201 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1261 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1321 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1381 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1441 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1501 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1561 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1621 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     1681 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     1741 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     1801 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1861 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     1921 gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt
     1981 atcatacaca tacgatttag gtgacactat agaacgcggc cgccagctga agcttcgtac
     2041 gctgcaggtc gacggatccc cgggttaatt aaggggatag cttatcctgg ttacttagac
     2101 tgttaaacta aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat
     2161 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     2221 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     2281 atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag
     2341 ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg
     2401 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     2461 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     2521 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     2581 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     2641 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     2701 gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa agactcacgt
     2761 ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2821 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     2881 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2941 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3001 tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat tccaggtatt
     3061 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3121 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3181 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3241 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     3301 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3361 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3421 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3481 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3541 tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     3601 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     3661 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     3721 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     3781 gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
     3841 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     3901 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3961 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4021 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4201 acagga
//
