LOCUS       sequence-287714-        4851 bp    DNA     circular SYN 15-APR-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 4851)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4851)
  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"
COMMENT     Sequence Label: pGBW-m4046533
FEATURES             Location/Qualifiers
     source          1..4851
                     /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"
     CDS             538..561
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(606..624)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             975..1778
                     /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      1861..2843
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2921..2950
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2972..3559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3571..3598)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3622..3813)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3814..4605,4606..4674))
                     /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     4437..4456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4675..4777)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4778..4809)
                     /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 aggttcttgg
      541 agccatccgc agtttgaaaa ataaacaggc cccttttcct ttgtcgatat catgtaatta
      601 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
      661 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
      721 gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc gccgaaccac
      781 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
      841 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
      901 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
      961 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     1021 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     1081 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     1141 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     1201 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     1261 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     1321 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     1381 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     1441 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     1501 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     1561 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     1621 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     1681 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     1741 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     1801 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     1861 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     1921 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     1981 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     2041 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     2101 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     2161 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     2221 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     2281 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     2341 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     2401 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     2461 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     2521 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     2581 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     2641 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     2701 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     2761 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     2821 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     2881 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     2941 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3061 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     3121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3421 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     3481 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     3541 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     3601 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3661 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3721 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3781 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     3841 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3901 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3961 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     4021 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4081 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4141 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4201 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4261 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4321 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     4381 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4441 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4501 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4561 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4621 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     4681 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     4741 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     4801 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag a
//
