LOCUS       sequence-297864-        6480 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 6480)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6480)
  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-m4046572
FEATURES             Location/Qualifiers
     source          1..6480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1075..1092
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2235..2253)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2604..3407
                     /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      3490..4472
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4550..4579
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4601..5188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5200..5227)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5251..5442)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5443..6234,6235..6303))
                     /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     6066..6085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6304..6406)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6407..6438)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gctatcgcca gtgaattttc ctcattacct agctatgcag ctttcgctac ggctcaggag
      541 gcctacgaac aagctgtagc caacggcgac tctgaagttg ttcttaagaa attgaagaag
      601 tctttgaatg tcgcgaaatc tgaatttgat agagatgctg ctatgcaacg taagctagaa
      661 aagatggctg accaagcaat gacccaaatg tacaaacaag ccagaagtga ggacaagcga
      721 gctaaggtga cttccgctat gcaaacaatg ctgttcacta tgttgagaaa gttggataac
      781 gatgctctga acaatattat aaacaatgcc agagatggtt gtgtccccct caacattatc
      841 ccattaacca ctgccgctaa attgatggtt gttattccgg attataatac atacaaaaac
      901 acttgcgacg gtaccacctt cacatacgcc tctgctcttt gggaaatcca acaggtagtc
      961 gacgctgact ccaagatagt tcaattgagc gaaatttcta tggataactc gccaaaccta
     1021 gcttggccat taatcgtcac tgcgttgaga gctaatagtg cggttaagtt gcaactcgtg
     1081 ccgagaggtt ctaaaatcga ggaaggtaag ctagtcattt ggataaatgg agataagggt
     1141 tacaacggtt tggcagaagt aggcaagaaa tttgaaaagg acacaggtat caaggtgacg
     1201 gttgaacatc cagacaagtt agaagaaaaa ttccctcaag ttgctgctac tggtgatggt
     1261 cccgacatta ttttctgggc ccacgatcgt tttggtggat atgctcagtc aggtttgctg
     1321 gctgagatca ccccagacaa ggccttccaa gataaattgt acccattcac ttgggatgct
     1381 gtcagataca acggtaagct tatagcatat ccaattgctg ttgaagcctt atcgttgatc
     1441 tacaataagg acttgttacc taacccacca aagacatggg aagaaattcc agctttggat
     1501 aaagagctaa aggccaaggg aaaaagcgct ttgatgttta acttacaaga accttatttc
     1561 acctggccat taatcgctgc tgacggtggt tacgcattta aatacgaaaa cggtaagtat
     1621 gacattaagg atgtcggcgt tgataatgct ggtgcaaagg ctggtttgac cttcttggtt
     1681 gatttaataa agaataaaca catgaacgct gacactgact actctatcgc cgaagccgct
     1741 ttcaacaagg gtgaaactgc tatgaccata aatggtccat gggcctggag taacattgat
     1801 acttctaaag taaactatgg tgtcactgtt ttgcctacat tcaaaggcca accatccaag
     1861 ccctttgtcg gtgttctatc cgctggcatc aatgctgctt ctccaaacaa ggagctagct
     1921 aaagaatttt tggaaaatta cttgttaact gacgaaggtt tggaggccgt taacaaggac
     1981 aagccattgg gtgctgtcgc tttgaaatct tacgaagaag aattggctaa ggatcccaga
     2041 attgccgcta ccatggagaa cgcccaaaag ggtgaaatta tgccaaacat cccacaaatg
     2101 tcagccttct ggtacgcagt taggactgcc gtgattaatg ctgcttccgg tagacaaacc
     2161 gtcgacgaag cattaaaaga tgctcaaacg taaacaggcc ccttttcctt tgtcgatatc
     2221 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     2281 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     2341 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     2401 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     2461 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     2521 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     2581 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     2641 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     2701 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     2761 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     2821 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     2881 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     2941 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     3001 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     3061 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     3121 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     3181 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     3241 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     3301 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     3361 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     3421 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     3481 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     3541 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     3601 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     3661 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     3721 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     3781 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     3841 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     3901 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     3961 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     4021 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     4081 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     4141 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     4201 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     4261 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     4321 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     4381 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     4441 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     4501 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     4561 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     4621 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4681 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     4741 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4801 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4861 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4921 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4981 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5041 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     5101 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     5161 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     5221 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     5281 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     5341 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5401 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     5641 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     6361 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     6421 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
