LOCUS       sequence-271036-        4191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal SRS tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4191)
  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 4191)
  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-SRS:kanMX6
FEATURES             Location/Qualifiers
     source          1..4191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(11..115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     183..201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(239..261)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     361..380
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        461..479
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     461..478
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      583..770
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(638..662)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            845..2201
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        845..1188
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1189..1998
                     /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(1256..1275)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2004..2201
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2305..2324)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2306..2324)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2411..2428)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2582..3170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2662..2681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3341..4132,4133..4191,1..10))
                     /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     3964..3983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
       61 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      121 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
      181 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
      241 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
      301 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
      361 cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg
      421 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
      481 cgcggccgcc agctgaagct tcgtacgctg caggtcgacg gatccccggg ttaattaact
      541 gcccgaaacc ggtggtggct aaggcgcgcc acttctaaat aagcgaattt cttatgattt
      601 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
      661 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
      721 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
      781 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
      841 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
      901 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
      961 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     1021 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     1081 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     1141 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa
     1201 aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1261 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1321 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1381 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1441 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca
     1501 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1561 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1621 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1681 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     1741 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     1801 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     1861 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     1921 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     1981 ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2041 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2101 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2161 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2221 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     2281 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2341 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2401 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2461 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2521 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2581 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2641 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2701 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2761 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2821 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2881 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2941 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3001 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3061 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3421 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3481 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3541 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3601 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3661 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3721 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3781 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3841 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3901 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3961 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4021 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4081 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4141 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt t
//
