LOCUS       sequence-298113-        5327 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5327)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5327)
  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-m4046477
FEATURES             Location/Qualifiers
     source          1..5327
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1002..1019
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1020..1037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1082..1100)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1451..2254
                     /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      2337..3319
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3397..3426
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3448..4035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4047..4074)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4098..4289)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4290..5081,5082..5150))
                     /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     4913..4932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5151..5253)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5254..5285)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg cacgtttctt tttcttgaaa attttttttt
      241 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa
      301 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
      361 ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaatggct atcgccagtg
      421 aattttcctc attacctagc tatgcagctt tcgctacggc tcaggaggcc tacgaacaag
      481 ctgtagccaa cggcgactct gaagttgttc ttaagaaatt gaagaagtct ttgaatgtcg
      541 cgaaatctga atttgataga gatgctgcta tgcaacgtaa gctagaaaag atggctgacc
      601 aagcaatgac ccaaatgtac aaacaagcca gaagtgagga caagcgagct aaggtgactt
      661 ccgctatgca aacaatgctg ttcactatgt tgagaaagtt ggataacgat gctctgaaca
      721 atattataaa caatgccaga gatggttgtg tccccctcaa cattatccca ttaaccactg
      781 ccgctaaatt gatggttgtt attccggatt ataatacata caaaaacact tgcgacggta
      841 ccaccttcac atacgcctct gctctttggg aaatccaaca ggtagtcgac gctgactcca
      901 agatagttca attgagcgaa atttctatgg ataactcgcc aaacctagct tggccattaa
      961 tcgtcactgc gttgagagct aatagtgcgg ttaagttgca actcgtgccg agaggttctc
     1021 accatcatca ccatcattaa acaggcccct tttcctttgt cgatatcatg taattagtta
     1081 tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta
     1141 gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac
     1201 gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt
     1261 gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga
     1321 agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga
     1381 aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg
     1441 aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt
     1501 tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga
     1561 tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt
     1621 actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct
     1681 aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat
     1741 tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga
     1801 cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc
     1861 agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg
     1921 ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga
     1981 ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg
     2041 gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta
     2101 tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact
     2161 atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga
     2221 agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg
     2281 tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg
     2341 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     2401 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     2461 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     2521 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     2581 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     2641 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     2701 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     2761 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     2821 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     2881 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     2941 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     3001 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     3061 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     3121 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     3181 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     3241 gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg
     3301 aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc
     3361 ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt
     3421 tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3481 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3541 aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3601 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3661 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3721 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3781 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3841 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3901 gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt
     3961 cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg
     4021 cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga
     4081 ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4141 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     4201 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     4261 ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc
     4321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4381 acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4441 tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt
     4501 gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc
     4741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4801 actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc
     4861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa
     4981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc
     5161 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     5221 tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt
     5281 tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
