LOCUS       sequence-453989-        4357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for yeast gene knockout, allows the use of 
            acetamide as sole nitrogen source.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4357)
  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 4357)
  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: SJZ9-amdS
FEATURES             Location/Qualifiers
     source          1..4357
                     /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(2607..2625)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2633..3221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2713..2732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3392..4183,4184..4252))
                     /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     4015..4034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4253..4357)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcattt aggtgacact atagagcggc cgccagctga agcctcgtac gctgctggtc
       61 gacgtattgg tgatggagca ggtttaatta accctaggtg aggcgcgcct agggataaca
      121 gggtaatcga cgtgcaccaa cttgcggaaa gtggaatccc gttccaaaac tggcatccac
      181 taattgatac atctacacac cgcacgcctt ttttctgaag cccactttcg tggactttgc
      241 catatgcaaa attcatgaag tgtgatacca agtcagcata cacctcacta gggtagtttc
      301 tttggttgta ttgatcattt ggttcatcgt ggttcattaa ttttttttct ccattgcttt
      361 ctggctttga tcttactatc atttggattt ttgtcgaagg ttgtagaatt gtatgtgaca
      421 agtggcacca agcatatata aaaaaaaaaa gcattatctt cctaccagag ttgattgtta
      481 aaaacgtatt tatagcaaac gcaattgtaa ttaattctta ttttgtatct tttcttccct
      541 tgtctcaatc ttttattttt attttatttt tcttttctta gtttctttca taacaccaag
      601 caactaatac tataacatac aataataatg ccacaatctt gggaagaatt ggctgctgac
      661 aagagagcta gattggctaa gactatccca gacgaatgga aggttcaaac tttgccagct
      721 gaagactctg ttatcgactt cccaaagaag tctggtatct tgtctgaagc tgaattgaag
      781 atcactgaag cttctgctgc tgacttggtt tctaagttgg ctgctggtga attgacttct
      841 gttgaagtta ctttggcttt ctgtaagaga gctgctatcg ctcaacaatt gactaactgt
      901 gctcacgaat tcttcccaga cgctgctttg gctcaagcta gagaattgga cgaatactac
      961 gctaagcaca agagaccagt tggtccattg cacggtttgc caatctcttt gaaggaccaa
     1021 ttgagagtta agggttacga aacttctatg ggttacatct cttggttgaa caagtacgac
     1081 gaaggtgact ctgttttgac tactatgttg agaaaggctg gtgctgtttt ctacgttaag
     1141 acttctgttc cacaaacttt gatggtttgt gaaactgtta acaacatcat cggtagaact
     1201 gttaacccaa gaaacaagaa ctggtcttgt ggtggttctt ctggtggtga aggtgctatc
     1261 gttggtatca gaggtggtgt tatcggtgtt ggtactgaca tcggtggttc tatcagagtt
     1321 ccagctgctt tcaacttctt gtacggtttg agaccatctc acggtagatt gccatacgct
     1381 aagatggcta actctatgga aggtcaagaa actgtacact ctgttgttgg tccaatcact
     1441 cactctgttg aagacttgag attgttcact aagtctgttt tgggtcaaga accatggaag
     1501 tacgactcta aggttatccc aatgccatgg agacaatctg aatctgacat catcgcttct
     1561 aagatcaaga acggtggttt gaacatcggt tactacaact tcgacggtaa cgttttgcca
     1621 cacccaccaa tcttgagagg tgttgaaact actgttgctg ctttggctaa ggctggtcac
     1681 actgttactc catggactcc atacaagcac gacttcggtc acgacttgat ctctcacatc
     1741 tacgctgctg acggttctgc tgacgttatg agagacatct ctgcttctgg tgaaccagct
     1801 atcccaaaca tcaaggactt gttgaaccca aacatcaagg ctgttaacat gaacgaattg
     1861 tgggacactc acttgcaaaa gtggaactac caaatggaat acttggaaaa gtggagagaa
     1921 gctgaagaaa aggctggtaa ggaattggac gctatcatcg ctccaatcac tccaactgct
     1981 gctgttagac acgaccaatt cagatactac ggttacgctt ctgttatcaa cttgttggac
     2041 ttcacttctg ttgttgttcc agttactttc gctgacaaga acatcgacaa gaagaacgaa
     2101 tctttcaagg ctgtttctga attggacgct ttggttcaag aagaatacga cccagaagct
     2161 taccacggtg ctccagttgc tgttcaagtt atcggtagaa gattgtctga agaaagaact
     2221 ttggctatcg ctgaagaagt tggtaagttg ttgggtaacg ttgttactcc ataaggagat
     2281 tgataagact tttctagttg catatctttt atatttaaat cttatctatt agttaatttt
     2341 ttgtaattta tccttatata tagtctggtt attctaaaat atcatttcag tatctaaaaa
     2401 ttcccctctt ttttcagtta tatcttaaca ggcgacagtc caaatgttga tttatcccag
     2461 tccgattcat cagggttgtg aagcattttg tcaatggtcg aaatcacatc agtaatagtg
     2521 cctcttactt gcctcataga atttctttct cttaacgtca ccgtttggtc ttttgagctc
     2581 gatccactag tggcctatgc ccgcggccta tagtgagtcg tattagttaa cttttccata
     2641 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2701 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2761 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2821 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2881 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2941 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3001 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3061 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3121 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3481 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3541 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3601 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3661 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3721 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3781 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3841 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3901 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3961 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4021 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4081 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4141 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4201 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4261 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4321 aatgtattta gaaaaataaa caaatagggg ttccgcg
//
