LOCUS       sequence-335170-        4845 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 4845)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4845)
  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-m4046521
FEATURES             Location/Qualifiers
     source          1..4845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             496..513
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             514..531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(576..594)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             945..1748
                     /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      1831..2813
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2891..2920
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2942..3529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3541..3568)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3592..3783)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3784..4575,4576..4644))
                     /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     4407..4426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4645..4747)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4748..4779)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
      361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaact ataatgtccg ctgacgcaca atcattccta
      481 aatgggtttg ctgtactcgt gccgagaggt tctcaccatc atcaccatca ttaaacaggc
      541 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
      601 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
      661 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
      721 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
      781 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
      841 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
      901 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
      961 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     1021 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     1081 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     1141 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     1201 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     1261 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     1321 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     1381 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     1441 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     1501 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     1561 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     1621 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     1681 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     1741 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     1801 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     1861 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     1921 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     1981 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     2041 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     2101 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     2161 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     2221 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     2281 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     2341 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     2401 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     2461 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     2521 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     2581 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     2641 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     2701 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     2761 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     2821 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     2881 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     2941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     3061 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3121 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3181 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3241 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3301 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3361 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3421 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3481 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3541 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     3601 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     3661 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     3721 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     3781 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3841 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     3901 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3961 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     4021 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4081 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4141 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4201 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     4261 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     4321 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4381 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4441 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     4501 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4561 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4621 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     4681 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     4741 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     4801 tgatttaaat ggtcagttag agcggccgct tgctatattg aagta
//
