LOCUS       sequence-298136-        5155 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5155)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5155)
  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-m4046469
FEATURES             Location/Qualifiers
     source          1..5155
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        10..405
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             782..799
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             800..865
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(910..928)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1279..2082
                     /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      2165..3147
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3225..3254
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3276..3863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3875..3902)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3926..4117)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4118..4909,4910..4978))
                     /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     4741..4760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4979..5081)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5082..5113)
                     /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 ataaaaattt ttatcacgtt tctttttctt
      241 gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      301 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
      361 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaat
      421 gaaattcttg gtttttcttg gtatcataac tacagtagcc gctttccacc aggaatgttc
      481 cttacaaagc tgcacgcaac atcaaccgta cgtcgttgat gacccatgtc caatccactt
      541 ctattcgaag tggtacatta gagttggagc taggaagtct gcacctttga ttgagctgtg
      601 tgtcgatgaa gctggttcaa agtctccaat tcaatacatc gacattggta attatactgt
      661 ctcctgtttg ccctttacca ttaactgcca agaaccaaaa ttaggcagtt tggttgtccg
      721 atgttctttc tacgaagact ttttggaata tcatgatgtg cgtgttgtcc tagatttcat
      781 cctcgtgccg agaggttctg attataaaga tcatgatggt gattataaag atcatgatat
      841 tgattataaa gatgatgatg ataaataaac aggccccttt tcctttgtcg atatcatgta
      901 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
      961 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     1021 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     1081 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     1141 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     1201 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     1261 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     1321 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     1381 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     1441 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     1501 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     1561 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     1621 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     1681 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     1741 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     1801 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     1861 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     1921 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     1981 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     2041 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     2101 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     2161 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     2221 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     2281 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     2341 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     2401 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     2461 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     2521 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     2581 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     2641 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     2701 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     2761 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     2821 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     2881 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     2941 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     3001 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     3061 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     3121 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     3181 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     3241 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3301 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3361 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     3421 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3481 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3541 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3601 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3661 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3721 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     3781 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     3841 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     3901 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     3961 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4021 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4081 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     4141 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4201 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4261 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     4321 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4381 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4441 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4501 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4561 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4621 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     4681 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4741 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4801 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4861 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4921 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     4981 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     5041 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     5101 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
