LOCUS       sequence-287655-        5925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5925)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5925)
  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"
COMMENT     Sequence Label: pGBW-m4046576
FEATURES             Location/Qualifiers
     source          1..5925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             520..537
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1680..1698)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2049..2852
                     /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"
     rep_origin      2935..3917
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3995..4024
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4046..4633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4645..4672)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4696..4887)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4888..5679,5680..5748))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5511..5530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5749..5851)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5852..5883)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tccgctgacg cacaatcatt cctaaatggg tttgctgtac tcgtgccgag aggttctaaa
      541 atcgaggaag gtaagctagt catttggata aatggagata agggttacaa cggtttggca
      601 gaagtaggca agaaatttga aaaggacaca ggtatcaagg tgacggttga acatccagac
      661 aagttagaag aaaaattccc tcaagttgct gctactggtg atggtcccga cattattttc
      721 tgggcccacg atcgttttgg tggatatgct cagtcaggtt tgctggctga gatcacccca
      781 gacaaggcct tccaagataa attgtaccca ttcacttggg atgctgtcag atacaacggt
      841 aagcttatag catatccaat tgctgttgaa gccttatcgt tgatctacaa taaggacttg
      901 ttacctaacc caccaaagac atgggaagaa attccagctt tggataaaga gctaaaggcc
      961 aagggaaaaa gcgctttgat gtttaactta caagaacctt atttcacctg gccattaatc
     1021 gctgctgacg gtggttacgc atttaaatac gaaaacggta agtatgacat taaggatgtc
     1081 ggcgttgata atgctggtgc aaaggctggt ttgaccttct tggttgattt aataaagaat
     1141 aaacacatga acgctgacac tgactactct atcgccgaag ccgctttcaa caagggtgaa
     1201 actgctatga ccataaatgg tccatgggcc tggagtaaca ttgatacttc taaagtaaac
     1261 tatggtgtca ctgttttgcc tacattcaaa ggccaaccat ccaagccctt tgtcggtgtt
     1321 ctatccgctg gcatcaatgc tgcttctcca aacaaggagc tagctaaaga atttttggaa
     1381 aattacttgt taactgacga aggtttggag gccgttaaca aggacaagcc attgggtgct
     1441 gtcgctttga aatcttacga agaagaattg gctaaggatc ccagaattgc cgctaccatg
     1501 gagaacgccc aaaagggtga aattatgcca aacatcccac aaatgtcagc cttctggtac
     1561 gcagttagga ctgccgtgat taatgctgct tccggtagac aaaccgtcga cgaagcatta
     1621 aaagatgctc aaacgtaaac aggccccttt tcctttgtcg atatcatgta attagttatg
     1681 tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga
     1741 caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt
     1801 tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc
     1861 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
     1921 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     1981 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
     2041 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     2101 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     2161 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     2221 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     2281 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     2341 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     2401 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     2461 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     2521 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     2581 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     2641 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     2701 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     2761 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     2821 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     2881 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa
     2941 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     3001 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     3061 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     3121 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     3181 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     3241 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     3301 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     3361 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     3421 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     3481 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     3541 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     3601 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     3661 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     3721 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     3781 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     3841 gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa
     3901 gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg ttacaagctt
     3961 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc
     4021 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4081 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4141 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4201 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4261 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4321 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4381 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4441 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4501 cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc tttatagtcc
     4561 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg
     4621 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt
     4681 acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     4741 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     4801 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     4861 aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg cacctatctc
     4921 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4981 gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5041 accggctcca gatttatcag caataaacca gccagccggg agggccgagc gcagaagtga
     5101 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5161 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5221 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5281 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgccag
     5341 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5401 tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5461 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5521 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact
     5581 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5641 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5701 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt tagcttcctt
     5761 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     5821 gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg
     5881 actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
