LOCUS       sequence-454027-        3284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, confers antibiotic 
            resistance to nourseothricin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3284)
  AUTHORS   Feng Y-H, Song J-Z, Zhu J, Xie Z
  TITLE     Template plasmids optimized for deletion of multiple genes in yeast 
            Saccharomyces cerevisiae.
  JOURNAL   Microbiol Spectr. 2024 Oct 10:e0132024. doi: 
            10.1128/spectrum.01320-24.
  PUBMED    39387598
REFERENCE   2  (bases 1 to 3284)
  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: "Microbiol 
            Spectr. 2024 Oct 10:e0132024. doi: 10.1128/spectrum.01320-24."
COMMENT     Sequence Label: SJZ4-NAT
FEATURES             Location/Qualifiers
     source          1..3284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7..24
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             629..1201
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(1534..1552)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1560..2148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1640..1659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2319..3110,3111..3179))
                     /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     2942..2961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3180..3284)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcatcgtac gccgccggtc
       61 gacgtattgg agacggcgca ggattaatta atcctaggtg aggcgcgcct agggataaca
      121 gggtaattgg gtcattacgt aaataatgat aggaatggga ttcttctatt tttccttttt
      181 ccattctagc agccgtcggg aaaacgtggc atcctctctt tcgggctcaa ttggagtcac
      241 gctgccgtga gcatcctctc tttccatatc taacaactga gcacgtaacc aatggaaaag
      301 catgagctta gcgttgctcc aaaaaagtat tggatggtta ataccatttg tctgttctct
      361 tctgactttg actcctcaaa aaaaaaaaaa tctacaatca acagatcgct tcaattacgc
      421 cctcacaaaa acttttttcc ttcttcttcg cccacgttaa attttatccc tcatgttgtc
      481 taacggattt ctgcacttga tttattataa aaagacaaag acataatact tctctatcaa
      541 tttcagttat tgttcttcct tgcgttattc ttctgttctt ctttttcttt tgtcatatat
      601 aaccataacc aagtaataca tattcaaaat gggtaccact cttgacgaca cggcttaccg
      661 gtaccgcacc agtgtcccgg gggacgccga ggccatcgag gcactggatg ggtccttcac
      721 caccgacacc gtcttccgcg tcaccgccac cggggacggc ttcaccctgc gggaggtgcc
      781 ggtggacccg cccctgacca aggtgttccc cgacgacgaa tcggacgacg aatcggacga
      841 cggggaggac ggcgacccgg actcccggac gttcgtcgcg tacggggacg acggcgacct
      901 ggcgggcttc gtggtcgtct cgtactccgg ctggaaccgc cggctgaccg tcgaggacat
      961 cgaggtcgcc ccggagcacc gggggcacgg ggtcgggcgc gcgttgatgg ggctcgcgac
     1021 ggagttcgcc cgcgagcggg gcgccgggca cctctggctg gaggtcacca acgtcaacgc
     1081 accggcgatc cacgcgtacc ggcggatggg gttcaccctc tgcggcctgg acaccgccct
     1141 gtacgacggc accgcctcgg acggcgagca ggcgctctac atgagcatgc cctgccccta
     1201 agtgaattta ctttaaatct tgcatttaaa taaattttct ttttatagct ttatgactta
     1261 gtttcaattt atatactatt ttaatgacat tttcgattca ttgattgaaa gctttgtgtt
     1321 ttttcttgat gcgctattgc attgttcttg tctttttcgc cacatgtaat atctgtagta
     1381 gatacctgat acattgtgga tgctgagtga aattttagtt aataatggag gcgctcttaa
     1441 taattttggg gatattggct ttttttttta aagtttacaa atgaattttt tccgccagga
     1501 tgagctcgat ccactagtgg cctatgcccg cggcctatag tgagtcgtat tagttaactt
     1561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcg
//
