LOCUS       sequence-300490-        5170 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 5170)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5170)
  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-m4046301
FEATURES             Location/Qualifiers
     source          1..5170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(286..1089)
                     /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"
     primer_bind     1440..1458
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(1503..1520)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1521..1538)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(1966..2379)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      2425..2456
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2457..2559
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2560..2628,2629..3420)
                     /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     complement(2778..2797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3421..3612
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      3636..3663
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3675..4262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4284..4313)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(4391..5170,1..203)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ttacaaaaat
       61 gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg ttctgtaaaa
      121 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttctaca
      181 aaatgaagca cagatgcttc gttactcggg taataactga tataattaaa ttgaagctct
      241 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
      301 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
      361 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagaaac
      421 cacatcatcc acggttctat actgttgacc caatgcatct cccttgtcat ctaaacccac
      481 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
      541 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
      601 ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
      661 ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
      721 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg
      781 caatttgact gtattaccaa tgtcagcaaa ttttctgtcc tcgaagagta aaaaattgta
      841 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat
      901 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
      961 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     1021 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     1081 tttcgacatg atttatcttc gtttcctgca cgtttttgtt ctgtgcagtt gggttaagaa
     1141 tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct
     1201 gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaagg
     1261 aaaccgtgta ctgcagtgca ctagtggttc ggcggccgct ttgtacagaa aaaaaagaaa
     1321 aatttgaaat ataaataacg ttcttaatac taacataact attaaaaaaa ataaataggg
     1381 acctagactt caggttgtct aactccttcc ttttcggtta gagcggatgt gggaggaggg
     1441 cgtgaatgta agcgtgacat aactaattac atgatatcga caaaggaaaa ggggcctgtt
     1501 taatgatggt gatgatggtg agaacctctc ggcacgagaa gcatgggttc gcggagttga
     1561 tcacaactac agccataacc tttccacata ccgcagacgg tacagactgt gtttttaagt
     1621 gtaaaaccca cagggtcatt agcacaagtt gtaggtattt gtacatactt accttttaag
     1681 tcacaaaatc ctttaggatt tggatgatct atgtggcaac ggcagtacag acaacacgat
     1741 gcaccaccaa aggattcttg atccatattg gcttccggtg taactgttat tgcctgacca
     1801 gtaccagtgt gtgtacacaa catcttaaca caattagtga ttggttgtcc cccactagct
     1861 agataatctt tgtaagcttt agcagcatct acagcaaaag cacagaaaga taatacagtt
     1921 gaattggcag gcacttctgt tgcattaccc attttgtaat taaaacttag attagattgc
     1981 tatgctttct ttctaatgag caagaagtaa aaaaagttgt aatagaacaa gaaaaatgaa
     2041 actgaaactt gagaaattga agaccgttta ttaacttaaa tatcaatggg aggtcatcga
     2101 aagagaaaaa aatcaaaaaa aaaaattttc aagaaaaaga aacgtgataa aaatttttat
     2161 tgcctttttc gacgaagaaa aagaaacgag gcggtctctt ttttcttttc caaaccttta
     2221 gtacgggtaa ttaacgacac cctagaggaa gaaagagggg aaatttagta tgctgtgctt
     2281 gggtgttttg aagtggtacg gcgatgcgcg gagtccgaga aaatctggaa gagtaaaaaa
     2341 ggagtagaaa cattttgaag ctatggtgtg tggggcggcc gctctaactg accatttaaa
     2401 tcatacctga cctccatagc agaaagtcaa aagcctccga ccggaggctt ttgacttgat
     2461 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     2521 tttgagttat cgagattttc aggagctaag gaagctaaaa tgagtattca acatttccgt
     2581 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2641 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2701 gatctcaaca gcggtaagat ccttgagagt ttacgccccg aagaacgttt tccaatgatg
     2761 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2821 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2881 gaaaagcatc tcacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2941 agtgataaca ctgcggccaa cttacttctg gcaacgatcg gaggaccgaa ggagctaacc
     3001 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3061 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3121 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3181 tggatggagg cggataaagt tgcaggatca cttctgcgct cggccctccc ggctggctgg
     3241 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3301 gggccagatg gtaagccctc ccgcatcgta gttatctaca cgacggggag tcaggcaact
     3361 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3421 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
     3481 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
     3541 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
     3601 gaaaacctct gatgagggcc caaatgtaat cacctggctc accttcgggt gggcctttct
     3661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgatgc
     3721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tacccctgga
     3781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4141 ctgaagccag ttacctcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4261 aagaagatcc tttgattttc taccgaagaa aggcccaccc gtgaaggtga gccagtgagt
     4321 tgattgcagt ccagttacgc tggagtcaag cttgtaacac gcggccgctc gggaagttcc
     4381 tattctctag ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg
     4441 atgacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc
     4501 gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat
     4561 atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct
     4621 ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac
     4681 tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat
     4741 agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta
     4801 tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc
     4861 ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga
     4921 gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac
     4981 ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa
     5041 aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca
     5101 aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg
     5161 taaaaatgca
//
