LOCUS       sequence-454020-        3343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, uses auxotrophic marker 
            TRP1 (Kluyveromyces lactis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3343)
  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 3343)
  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: SJZ2-TRP1
FEATURES             Location/Qualifiers
     source          1..3343
                     /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"
     promoter        complement(1593..1611)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1619..2207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1699..1718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2378..3169,3170..3238))
                     /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     3001..3020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3239..3343)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcttcgtac gcagcaggtc
       61 gacggatcgg tgacggagct ggcttaatta atcctaggtg aggcgcgcct agggataaca
      121 gggtaatttt gagttattca tttccagcac ttccaggata aaagttacta ccaactgaaa
      181 acaagagaaa ccgccgtgaa actatggagc actacagtta aaacaccact aactccactc
      241 tgttgacttt tatctgtaac tcctcatgac aaccttgttg ataattatct caagatgacg
      301 atatttacat tgtcattata ttccattttc gcgtctcata tgttacccgt atcaaagagt
      361 ataaccctat caataagtga tgtgtatcat ttataaacct catattttaa gaataaacct
      421 atataatata ggaatatacc ctgaaaatag ttagaggtct acaatgcccc tattttacct
      481 ccagaattat aatctcctta agacttcaac tagaagtggt atttactagc acactgtgaa
      541 acagtagcgt caagtcggta gtaaacatca atcgcgtccc aactgcaaga gctactagaa
      601 gtaaacacag tcttacccaa actaaggatg ctcgttaaag tgtgtggttt gcaaaccgtt
      661 gaagctgcaa agactgctgt ggatgatggt gctgattact taggtatcat ttgtgttccc
      721 ggtaggaaaa gaaccattga ctcatctgtt gcgaaaggta tttcaactgc agttcaccaa
      781 caagagaacg tgaaaggtac taaactagtc ggggtgttta gaaatcagtc cgttgatgat
      841 gtccttcaac tgtaccacga atataatcta gatgtgatac aattacatgg agatgaagat
      901 attaaagaat acagatcttt gatcccatct tcaattccaa tcattaagag gttccagttc
      961 ccacaggatt gtgaattact actggacctg tatgaacacg tagacaatgt gctgacgttg
     1021 ttcgattctg gtgaaggtgg cactggtgag aaattgaatt ggagtgcaat ttccagttgg
     1081 tctgcaagtc atcccgagat aaaattcatt atcgctggtg gattgaatcc tgataacgtt
     1141 tctgttgcca ttaatatgtt accaaatgcg atcggtgtcg atgtaagtgg aggagtagag
     1201 actgatggta tcaaggattt agaaaaggta aagctattca tccagcaggc ctctcaatag
     1261 gcttagcgat cgccctgtaa aatatccgaa aagttttact tttgctttcc tctgtatttt
     1321 ttaatttttt tacgataaac taaatcatat aatattttat atagaaatga taaaaaaaga
     1381 ataagaacaa taataaaaaa atcaataata ttattcaatt gtacttatga ataaaaaaaa
     1441 aaataatagt gacttgaaga ctaggaatat cggtcatttt cccctataaa taaaaaataa
     1501 aaagagctct ttaataataa aaacttgata atatcaacag tatacaatgt gtatatacgc
     1561 gagctcgatc cactagtggc ctatgcccgc ggcctatagt gagtcgtatt agttaacttt
     1621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2041 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2221 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2281 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2341 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2401 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2461 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2521 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2581 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2641 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2701 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2761 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2821 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2881 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2941 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3001 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3061 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3121 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3181 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3241 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3301 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcg
//
