LOCUS       sequence-271093-        4206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal A1 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-A1:kanMX6
FEATURES             Location/Qualifiers
     source          1..4206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      13..200
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(68..92)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            275..1631
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        275..618
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             619..1428
                     /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(686..705)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1434..1631
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1735..1754)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1736..1754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1841..1858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2012..2600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2092..2111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2771..3562,3563..3631))
                     /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     3394..3413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3632..3736)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3804..3822
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3860..3882)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3982..4001
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4082..4100
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4082..4099
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
       61 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
      121 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt
      181 attgaccaca cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta
      241 gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct
      301 tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc
      361 catacatccc catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag
      421 caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc
      481 gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact
      541 gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac
      601 atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc gcgattaaat
      661 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
      721 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
      781 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
      841 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
      901 ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata tcctgattca
      961 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1021 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1081 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1141 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     1201 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1261 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1321 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     1381 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agtactgaca
     1441 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     1501 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     1561 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     1621 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtttaaacg agctcgaatt
     1681 catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     1741 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     1801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3781 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3841 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3901 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3961 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
     4021 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     4081 gatttaggtg acactataga acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac
     4141 ggatccccgg gttaattaag gggatagcct ggacatgctc gagtggtctc tgatgtaagg
     4201 cgcgcc
//
