LOCUS       sequence-305752-        6438 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 6438)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6438)
  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-m4046256
FEATURES             Location/Qualifiers
     source          1..6438
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      1014..1043
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1065..1652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1664..1691)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1715..1906)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1907..2698,2699..2767))
                     /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     2530..2549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2768..2870)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2871..2902)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2948..3361
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3972..3989
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(5132..5150)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             5501..6304
                     /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"
ORIGIN
        1 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
       61 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
      121 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
      181 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
      241 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
      301 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
      361 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
      421 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
      481 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
      541 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
      601 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
      661 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
      721 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
      781 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
      841 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg
      901 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc
      961 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac
     1021 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     1081 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1141 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     1201 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1261 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1321 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     1381 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     1441 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     1501 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg
     1561 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     1621 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     1681 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     1741 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     1801 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     1861 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     1921 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1981 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     2041 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     2101 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     2161 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2221 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2281 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2341 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2401 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     2461 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2521 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2581 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2641 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2701 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2761 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     2821 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     2881 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt
     2941 tagagcggcc gccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc
     3001 agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact
     3061 aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa
     3121 agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat
     3181 cacgtttctt tttcttgaaa attttttttt ttgatttttt tctctttcga tgacctccca
     3241 ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt
     3301 tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa
     3361 gttttaatta caaaatggct atcgccagtg aattttcctc attacctagc tatgcagctt
     3421 tcgctacggc tcaggaggcc tacgaacaag ctgtagccaa cggcgactct gaagttgttc
     3481 ttaagaaatt gaagaagtct ttgaatgtcg cgaaatctga atttgataga gatgctgcta
     3541 tgcaacgtaa gctagaaaag atggctgacc aagcaatgac ccaaatgtac aaacaagcca
     3601 gaagtgagga caagcgagct aaggtgactt ccgctatgca aacaatgctg ttcactatgt
     3661 tgagaaagtt ggataacgat gctctgaaca atattataaa caatgccaga gatggttgtg
     3721 tccccctcaa cattatccca ttaaccactg ccgctaaatt gatggttgtt attccggatt
     3781 ataatacata caaaaacact tgcgacggta ccaccttcac atacgcctct gctctttggg
     3841 aaatccaaca ggtagtcgac gctgactcca agatagttca attgagcgaa atttctatgg
     3901 ataactcgcc aaacctagct tggccattaa tcgtcactgc gttgagagct aatagtgcgg
     3961 ttaagttgca actcgtgccg agaggttcta aaatcgagga aggtaagcta gtcatttgga
     4021 taaatggaga taagggttac aacggtttgg cagaagtagg caagaaattt gaaaaggaca
     4081 caggtatcaa ggtgacggtt gaacatccag acaagttaga agaaaaattc cctcaagttg
     4141 ctgctactgg tgatggtccc gacattattt tctgggccca cgatcgtttt ggtggatatg
     4201 ctcagtcagg tttgctggct gagatcaccc cagacaaggc cttccaagat aaattgtacc
     4261 cattcacttg ggatgctgtc agatacaacg gtaagcttat agcatatcca attgctgttg
     4321 aagccttatc gttgatctac aataaggact tgttacctaa cccaccaaag acatgggaag
     4381 aaattccagc tttggataaa gagctaaagg ccaagggaaa aagcgctttg atgtttaact
     4441 tacaagaacc ttatttcacc tggccattaa tcgctgctga cggtggttac gcatttaaat
     4501 acgaaaacgg taagtatgac attaaggatg tcggcgttga taatgctggt gcaaaggctg
     4561 gtttgacctt cttggttgat ttaataaaga ataaacacat gaacgctgac actgactact
     4621 ctatcgccga agccgctttc aacaagggtg aaactgctat gaccataaat ggtccatggg
     4681 cctggagtaa cattgatact tctaaagtaa actatggtgt cactgttttg cctacattca
     4741 aaggccaacc atccaagccc tttgtcggtg ttctatccgc tggcatcaat gctgcttctc
     4801 caaacaagga gctagctaaa gaatttttgg aaaattactt gttaactgac gaaggtttgg
     4861 aggccgttaa caaggacaag ccattgggtg ctgtcgcttt gaaatcttac gaagaagaat
     4921 tggctaagga tcccagaatt gccgctacca tggagaacgc ccaaaagggt gaaattatgc
     4981 caaacatccc acaaatgtca gccttctggt acgcagttag gactgccgtg attaatgctg
     5041 cttccggtag acaaaccgtc gacgaagcat taaaagatgc tcaaacgtaa acaggcccct
     5101 tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca
     5161 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     5221 ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     5281 tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt
     5341 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     5401 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     5461 actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     5521 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     5581 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     5641 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     5701 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     5761 actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     5821 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     5881 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     5941 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     6001 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     6061 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     6121 tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg
     6181 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     6241 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     6301 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     6361 ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa
     6421 aatgcaacgc gagagcgc
//
