LOCUS       sequence-327694-        5183 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Knocking out GAL80 using KlURA3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5183)
  AUTHORS   Lu Z, Peng B, Ebert BE, Dumsday G, Vickers CE
  TITLE     Auxin-mediated protein depletion for metabolic engineering in 
            terpene-producing yeast.
  JOURNAL   Nat Commun. 2021 Feb 16;12(1):1051. doi: 10.1038/s41467-021-21313-1.
  PUBMED    33594068
REFERENCE   2  (bases 1 to 5183)
  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: "Nat 
            Commun."; date: "2021-02-16"; volume: "12(1):1051. doi"; pages: " 
            10.1038/s41467-021-21313-1"
COMMENT     Sequence Label: pIR3DH8
FEATURES             Location/Qualifiers
     source          1..5183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..41)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(25..41)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(38..60)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    49..65
                     /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(73..79,80..97,98..103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    118..139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(256..273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(427..1015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(507..526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1186..1977,1978..2046))
                     /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     1809..1828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2047..2151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2219..2237
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2275..2297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2397..2416
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2610..2632
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2624..2641
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2625..2641
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3217..3432
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3433..4236
                     /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 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
       61 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2221 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     2281 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     2341 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     2401 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     2461 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     2521 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     2581 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     2641 tgaattcgtt taaacggccg cctctgccat ggcaaagaat gctttccatg acgatcatcg
     2701 tagtgcccaa ttgggtgcct ctatgatggg tatggcttgg gcaagtgtct ttttatgtat
     2761 cgtggaattt atcctgctgg tcttctggtc tgttagggca aggttggcct ctacttactc
     2821 catcgacaat tcaagataca gaacctcctc cagatggaat cccttccata gagagaagga
     2881 gcaagcaact gacccaatat tgactgccac tggacctgaa gacatgcaac aaagtgcaag
     2941 catagtgggg ccttcttcca atgctaatcc ggtcactgcc actgctgcta cggaaaacca
     3001 acctaaaggt attaacttct tcactataag aaaatcacac gagcgcccgg acgatgtctc
     3061 tgtttaaaga tgctaagaga tagtgatgat atttcataaa taatgtaatt ctatatatgt
     3121 taattacctt ttttgcgagg catatttatg gtgaaggata agttttgacc atcaaagaag
     3181 gttaatgtgg ctgtggtttc agggtccata aagcttttca attcatcatt ttttttttat
     3241 tctttttttt gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     3301 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     3361 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa
     3421 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     3481 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     3541 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     3601 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     3661 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac
     3721 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     3781 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     3841 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     3901 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     3961 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     4021 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag
     4081 tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga
     4141 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     4201 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     4261 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgggaat
     4321 ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa ttggaaagaa aaagcttcat
     4381 ggcctttata aaaaggaact atccaatacc tcgccagaac caagtaacag tattttacgg
     4441 ggcacaaatc aagaacaata agacaggact gtaaagctcg aggactacaa caagagatct
     4501 tcggtctcaa ccgtgcctaa tgcagctccc ataagagtcg gattcgtcgg tctcaacgca
     4561 gccaaaggat gggcaatcaa gacacattac cccgccatac tgcaactatc gtcacaattt
     4621 caaatcactg ccttatacag tccaaaaatt gagacttcta ttgccaccat ccagcgtcta
     4681 aaattgagta atgccactgc ttttcccact ttagagtcat ttgcatcatc ttccactata
     4741 gatatgatag tgatagctat ccaagtggcc agtcattatg acgttgttat gcctctcttg
     4801 gaattctcca aaaataatcc gaacctcaag tatcttttcg tagaatgggc ccttgcatgt
     4861 tcactagatc aagccgaatc catttataag gctgctgctg aacgtggggt tcaaaccatc
     4921 atctctttac aaggtcgtaa atcaccatat attttgagag caaaagaatt aatatctcaa
     4981 ggctatatcg gcgacattaa ttctatcgag attgctggaa atggcggttg gtacggctac
     5041 gaaaggcctg ttaaatcacc aaaatacatc tatgaaatcg ggaacggtgt agatctggta
     5101 accacaacat ttggtcacac aatcgatatt ttacaataca tgacaagttc gtacttttcc
     5161 aggataaatg caatggttta aac
//
