LOCUS       sequence-287629-        5347 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 5347)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5347)
  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-m4046417
FEATURES             Location/Qualifiers
     source          1..5347
                     /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             1022..1039
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1040..1057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1102..1120)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1471..2274
                     /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      2357..3339
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3417..3446
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3468..4055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4067..4094)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4118..4309)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4310..5101,5102..5170))
                     /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     4933..4952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5171..5273)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5274..5305)
                     /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 atgatagcct cagagtttag ttcccttcca tcatatgcag cttttgctac
      481 tgctcaagaa gcttatgagc aggctgttgc taatggtgat tctgaagttg ttcttaaaaa
      541 gttgaagaag tctttgaatg tggctaaatc tgaatttgac cgtgatgcag ccatgcaacg
      601 taagttggaa aagatggctg atcaagctat gacccaaatg tataaacagg ctagatctga
      661 ggacaagagg gcaaaagtta ctagtgctat gcagacaatg cttttcacta tgcttagaaa
      721 gttggataat gatgcactca acaacattat caacaatgca agagatggtt gtgttccctt
      781 gaacataata cctcttacaa cagcagccaa actaatggtt gtcataccag actataacac
      841 atataaaaat acgtgtgatg gtacaacatt tacttatgca tcagcattgt gggaaatcca
      901 acaggttgta gatgcagata gtaaaattgt tcaacttagt gaaattagta tggacaattc
      961 acctaattta gcatggcctc ttattgtaac agctttaagg gccaattctg ctgtcaaatt
     1021 actcgtgccg agaggttctc accatcatca ccatcattaa acaggcccct tttcctttgt
     1081 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     1141 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     1201 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     1261 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     1321 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1381 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1441 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1501 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1561 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     1621 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     1681 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     1741 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     1801 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     1861 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     1921 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     1981 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     2041 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     2101 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     2161 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     2221 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     2281 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     2341 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2401 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2461 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2521 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2581 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2641 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     2701 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     2761 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     2821 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     2881 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     2941 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     3001 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     3061 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     3121 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     3181 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     3241 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     3301 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     3361 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     3421 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     3481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3541 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     3601 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3661 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3721 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3781 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3841 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3901 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     3961 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     4021 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     4081 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     4141 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4201 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4261 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     4321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4381 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     4441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4501 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     4561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4741 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4801 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     4861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4981 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5161 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     5221 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     5281 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     5341 agttaga
//
