LOCUS       sequence-287663-        5229 bp    DNA     circular SYN 15-DEC-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 5229)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5229)
  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-m4046583
FEATURES             Location/Qualifiers
     source          1..5229
                     /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"
     CDS             916..939
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(984..1002)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1353..2156
                     /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      2239..3221
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3299..3328
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3350..3937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3949..3976)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4000..4191)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4192..4983,4984..5052))
                     /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     4815..4834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5053..5155)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5156..5187)
                     /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 gttcttggag ccatccgcag tttgaaaaat aaacaggccc cttttccttt
      961 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     1021 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     1081 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1141 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     1201 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1261 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     1321 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     1381 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     1441 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     1501 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     1561 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     1621 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     1681 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     1741 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     1801 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     1861 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     1921 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     1981 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     2041 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     2101 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     2161 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     2221 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     2281 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2341 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     2401 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2461 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     2521 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     2581 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     2641 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     2701 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     2761 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     2821 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     2881 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     2941 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     3001 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     3061 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     3121 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     3181 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     3241 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     3301 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3421 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3481 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3541 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3601 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3661 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3721 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3781 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     3841 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     3901 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     3961 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     4021 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     4081 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     4141 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     4201 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4261 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     4321 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4381 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     4441 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4501 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4561 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4621 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     4681 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     4741 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4801 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4861 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4921 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4981 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5041 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     5101 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     5161 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     5221 tcagttaga
//
