LOCUS       sequence-291467-        6303 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6303)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6303)
  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-m4046510
FEATURES             Location/Qualifiers
     source          1..6303
                     /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             898..915
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2058..2076)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2427..3230
                     /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      3313..4295
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4373..4402
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4424..5011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5023..5050)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5074..5265)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5266..6057,6058..6126))
                     /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     5889..5908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6127..6229)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6230..6261)
                     /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 gcaggaaacg ctactgaagt tccagccaat tcaaccgtgt tgagcttttg tgctttcgcg
      541 gtagatgctg ccaaggctta caaagactat cttgcttccg gtgggcaacc cattacgaac
      601 tgcgtcaaga tgctgtgtac tcacacaggc accggtcagg ctataactgt taccccagag
      661 gccaatatgg accaagaatc gttcggtggt gcatcttgtt gtctctactg ccgttgtcat
      721 atcgatcacc caaaccctaa gggtttttgt gacttaaaag gtaagtacgt tcaaatccca
      781 actacctgtg ctaacgatcc agtcggtttc accttgaaga atacagtttg tactgtatgc
      841 ggtatgtgga aaggatacgg ttgttcttgc gatcaattga gagaacctat gttgcaactc
      901 gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa tggagataag
      961 ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg tatcaaggtg
     1021 acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc tactggtgat
     1081 ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca gtcaggtttg
     1141 ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt cacttgggat
     1201 gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc cttatcgttg
     1261 atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat tccagctttg
     1321 gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca agaaccttat
     1381 ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga aaacggtaag
     1441 tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt gaccttcttg
     1501 gttgatttaa taaagaataa acacatgaac gctgacactg actactctat cgccgaagcc
     1561 gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg gagtaacatt
     1621 gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg ccaaccatcc
     1681 aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa caaggagcta
     1741 gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc cgttaacaag
     1801 gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc taaggatccc
     1861 agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa catcccacaa
     1921 atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc cggtagacaa
     1981 accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc ctttgtcgat
     2041 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     2101 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     2161 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     2221 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     2281 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     2341 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     2401 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     2461 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     2521 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     2581 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     2641 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     2701 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     2761 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     2821 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     2881 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     2941 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     3001 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     3061 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     3121 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     3181 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     3241 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     3301 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     3361 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3421 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     3481 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3541 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     3601 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     3661 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     3721 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     3781 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     3841 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     3901 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     3961 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     4021 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     4081 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     4141 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     4201 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     4261 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     4321 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     4381 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     4441 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4501 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     4561 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4621 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4681 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4741 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4801 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4861 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     4921 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     4981 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     5041 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     5101 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     5161 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     5221 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     5281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5341 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     5401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     5461 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5701 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5761 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     5821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5941 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6121 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     6181 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     6241 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     6301 aga
//
