LOCUS       sequence-271064-        3957 bp    DNA     circular SYN 15-APR-2025
DEFINITION  PCR-based C-terminal SRS tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3957)
  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 3957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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:natMX4
FEATURES             Location/Qualifiers
     source          1..3957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        189..207
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     189..206
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      311..498
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(366..390)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            573..1692
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        573..916
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             917..1489
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1495..1692
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1799..1818)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1800..1818)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1905..1922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2076..2664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2156..2175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2835..3626,3627..3695))
                     /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     3458..3477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3696..3800)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3868..3886
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3924..3946)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
       61 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
      121 catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata
      181 cacatacgat ttaggtgaca ctatagaacg cggccgccag ctgaagcttc gtacgctgca
      241 ggtcgacgga tccccgggtt aattaactgc ccgaaaccgg tggtggctaa ggcgcgccac
      301 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
      361 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
      421 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
      481 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc
      541 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
      601 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
      661 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
      721 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
      781 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
      841 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
      901 ccgaacataa acaaccatgg gtaccactct tgacgacacg gcttaccggt accgcaccag
      961 tgtcccgggg gacgccgagg ccatcgaggc actggatggg tccttcacca ccgacaccgt
     1021 cttccgcgtc accgccaccg gggacggctt caccctgcgg gaggtgccgg tggacccgcc
     1081 cctgaccaag gtgttccccg acgacgaatc ggacgacgaa tcggacgacg gggaggacgg
     1141 cgacccggac tcccggacgt tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt
     1201 ggtcgtctcg tactccggct ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc
     1261 ggagcaccgg gggcacgggg tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg
     1321 cgagcggggc gccgggcacc tctggctgga ggtcaccaac gtcaacgcac cggcgatcca
     1381 cgcgtaccgg cggatggggt tcaccctctg cggcctggac accgccctgt acgacggcac
     1441 cgcctcggac ggcgagcagg cgctctacat gagcatgccc tgcccctaat cagtactgac
     1501 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     1561 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     1621 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     1681 gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa aacgagctcg
     1741 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     1801 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     1861 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2701 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2761 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2821 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2941 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3421 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3601 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3661 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3721 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3781 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     3841 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     3901 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcaca
//
