LOCUS       sequence-361230-        5970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S. cerevisiae Sigma1278b AMN1 gene knock out plasmid with KanMX 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5970)
  AUTHORS   Guinn L, Lo E, Balazsi G
  TITLE     Drug-dependent growth curve reshaping reveals mechanisms of 
            antifungal resistance in Saccharomyces cerevisiae.
  JOURNAL   Commun Biol. 2022 Mar 31;5(1):292. doi: 10.1038/s42003-022-03228-9.
  PUBMED    35361876
REFERENCE   2  (bases 1 to 5970)
  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: "Commun 
            Biol."; date: "2022-03-31"; volume: "5(1):292. doi"; pages: " 
            10.1038/s42003-022-03228-9"
COMMENT     Sequence Label: pBY011-AMN1ko
FEATURES             Location/Qualifiers
     source          1..5970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            505..1861
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        505..848
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             849..1658
                     /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(916..935)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1664..1861
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2389..2405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2389..2405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2402..2424)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2413..2429
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2437..2443,2444..2461,2462..2467))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2482..2503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2620..2637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2791..3379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2871..2890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3550..4341,4342..4410))
                     /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     4173..4192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4411..4515)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4583..4601
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4626..4841
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4842..5645
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 gaactaacaa aaacgttcaa aaagttttaa aactatatgt gttaactgta caaaaataac
       61 ttatttatca tatcattttt ttctctgttt atttcttcta gaacttatac ctgtcttttc
      121 cttttattct ttgaatttgg tttaatatcc ctttttggtt taatatccat ccattccttt
      181 cacttagaac taataattcc cttcgtttga taatttatca ttttcctttt ctgttagtaa
      241 agtacccatt aaatgaaact agaacgcgta agcagcaatg gctccttcaa aagaggtaga
      301 gatatccagt cacttgaatc tccgtgtact agaccgctga aaaaaatgtc acctagtcca
      361 tctttcacat ctcttaaaat ggagaagcct ttcaaagata ttgtaaggaa gtatggaggt
      421 catcttcatc aaagtagtta caatcccggc tcaagcaaag tggagttagt aagaccagat
      481 ttgtctttaa aaacagatca aagtgacatg gaggcccaga ataccctcct tgacagtctt
      541 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
      601 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
      661 ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
      721 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
      781 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
      841 aaacaaccat gggtaaggaa aagactcacg tttcgaggcc gcgattaaat tccaacatgg
      901 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
      961 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1021 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1081 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     1141 cgatccccgg caaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     1201 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     1261 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     1321 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     1381 aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     1441 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     1501 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     1561 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     1621 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agtactgaca ataaaaagat
     1681 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     1741 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     1801 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     1861 gaactcattt ccaaatctgg ctgttttaga gatcagaaat ttagataaaa taactgatgt
     1921 gaggcatttc gtgaagtaca acttgtggaa aaaatcattg gatgccccca ttctaatcga
     1981 agcatgtgaa cgtattacga aactaatcga tcaagaagaa aatagagtta agaggattaa
     2041 ctccttggtt gccttaaaag atatgacagc ttgggttaat gcagacgacg aaatagagaa
     2101 taatgtggac tagatggttt ttgtgacttt acctataaat agtacacaac agaccaccag
     2161 taattctaca cacttcttaa ctgataatat tattataatt gtaacttttt agcagcacta
     2221 aatttaatga atacatagat ttttaactag cattttacta ttctgtactt tttacttgaa
     2281 attccagaag ggccgaagaa accagaattc cttcacagaa aactattgaa ctacagaatt
     2341 ctccaaaaga tacactcata gtgttcatgc aagcttggcg taatcatggt catagctgtt
     2401 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2461 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2521 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2581 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2641 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2701 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3601 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3961 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4561 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt
     4621 agcttttcaa ttcatcattt tttttttatt cttttttttg atttcggttt ccttgaaatt
     4681 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     4741 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     4801 aactgcacag aacaaaaaca tgcaggaaac gaagataaat catgtcgaaa gctacatata
     4861 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     4921 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     4981 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     5041 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     5101 tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     5161 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     5221 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     5281 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     5341 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     5401 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     5461 atgacaaggg agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag
     5521 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     5581 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     5641 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     5701 tttaattata tcagttatta cccaattctc atgtttgaca gcttatcatc gatcgtccaa
     5761 ctgcatggag atgagtcgtg gcaagaatac caagagttcc tcggtttgcc agttattaaa
     5821 agactcgtat ttccaaaaga ctgcaacata ctactcagtg cagcttcaca gaaacctcat
     5881 tcgtttattc ccttgtttga ttcagaagca ggtgggacag gtgaactttt ggattggaac
     5941 tcgatttctg actgggttgg aaggcaagag
//
