LOCUS       sequence-287682-        4887 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 4887)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4887)
  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-m4046530
FEATURES             Location/Qualifiers
     source          1..4887
                     /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             514..531
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             532..597
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(642..660)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1011..1814
                     /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      1897..2879
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2957..2986
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3008..3595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3607..3634)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3658..3849)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3850..4641,4642..4710))
                     /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     4473..4492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4711..4813)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4814..4845)
                     /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 gctgatgcac aatcgttttt aaacgggttt gcgctcgtgc cgagaggttc tgattataaa
      541 gatcatgatg gtgattataa agatcatgat attgattata aagatgatga tgataaataa
      601 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
      661 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
      721 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
      781 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
      841 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      901 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      961 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     1021 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     1081 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     1141 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     1201 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     1261 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     1321 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     1381 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     1441 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     1501 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     1561 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1621 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1681 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1741 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1801 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1861 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     1921 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     1981 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     2041 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     2101 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     2161 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     2221 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     2281 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     2341 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     2401 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     2461 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     2521 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     2581 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     2641 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     2701 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     2761 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     2821 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     2881 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     2941 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     3001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     3121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3481 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3541 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3601 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     3661 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3721 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3781 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3841 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4021 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     4081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     4321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     4381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4501 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     4561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4681 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     4741 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     4801 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     4861 caggtatgat ttaaatggtc agttaga
//
