LOCUS       sequence-291570-        5359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5359)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5359)
  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-m4046269
FEATURES             Location/Qualifiers
     source          1..5359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1028..1045
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1046..1069
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1114..1132)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1483..2286
                     /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      2369..3351
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3429..3458
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3480..4067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4079..4106)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4130..4321)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4322..5113,5114..5182))
                     /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     4945..4964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5183..5285)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5286..5317)
                     /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 tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggctatcg ccagtgaatt ttcctcatta cctagctatg cagctttcgc
      481 tacggctcag gaggcctacg aacaagctgt agccaacggc gactctgaag ttgttcttaa
      541 gaaattgaag aagtctttga atgtcgcgaa atctgaattt gatagagatg ctgctatgca
      601 acgtaagcta gaaaagatgg ctgaccaagc aatgacccaa atgtacaaac aagccagaag
      661 tgaggacaag cgagctaagg tgacttccgc tatgcaaaca atgctgttca ctatgttgag
      721 aaagttggat aacgatgctc tgaacaatat tataaacaat gccagagatg gttgtgtccc
      781 cctcaacatt atcccattaa ccactgccgc taaattgatg gttgttattc cggattataa
      841 tacatacaaa aacacttgcg acggtaccac cttcacatac gcctctgctc tttgggaaat
      901 ccaacaggta gtcgacgctg actccaagat agttcaattg agcgaaattt ctatggataa
      961 ctcgccaaac ctagcttggc cattaatcgt cactgcgttg agagctaata gtgcggttaa
     1021 gttgcaactc gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc
     1081 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     1141 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     1201 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1261 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     1321 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     1381 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     1441 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     1501 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     1561 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     1621 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     1681 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     1741 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     1801 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     1861 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     1921 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     1981 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     2041 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     2101 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     2161 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     2221 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     2281 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     2341 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     2401 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2461 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     2521 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2581 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     2641 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     2701 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     2761 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     2821 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     2881 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     2941 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     3001 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     3061 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     3121 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     3181 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     3241 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     3301 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     3361 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     3421 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     3481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     3601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3961 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4021 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4081 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     4141 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4201 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4261 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4321 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4381 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     4441 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4501 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     4561 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4621 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4681 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4741 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     4801 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     4861 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4921 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4981 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     5041 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5101 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5161 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5221 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5281 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     5341 atttaaatgg tcagttaga
//
