LOCUS       sequence-307336-        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5083)
  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-m4046611
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(537..555)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             906..1709
                     /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      1792..2774
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2852..2881
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2903..3490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3502..3529)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3553..3744)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3745..4536,4537..4605))
                     /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     4368..4387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4606..4708)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4709..4740)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        join(4786..5083,1..116)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat
       61 tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa tctaagtttt
      121 aattacaaaa tgaaattctt ggtttttctt ggtatcataa ctacagtagc cgctttccac
      181 caggaatgtt ccttacaaag ctgcacgcaa catcaaccgt acgtcgttga tgacccatgt
      241 ccaatccact tctattcgaa gtggtacatt agagttggag ctaggaagtc tgcacctttg
      301 attgagctgt gtgtcgatga agctggttca aagtctccaa ttcaatacat cgacattggt
      361 aattatactg tctcctgttt gccctttacc attaactgcc aagaaccaaa attaggcagt
      421 ttggttgtcc gatgttcttt ctacgaagac tttttggaat atcatgatgt gcgtgttgtc
      481 ctagatttca tctaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
      541 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
      601 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
      661 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
      721 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
      781 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
      841 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
      901 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
      961 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     1021 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     1081 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     1141 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     1201 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     1261 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     1321 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     1381 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     1441 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     1501 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     1561 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     1621 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     1681 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     1741 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     1801 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     1861 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     1921 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     1981 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     2041 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     2101 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     2161 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     2221 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     2281 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     2341 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     2401 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     2461 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     2521 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     2581 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     2641 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     2701 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     2761 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     2821 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     2881 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2941 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3001 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3061 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3121 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3181 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3241 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3301 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3361 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     3421 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     3481 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     3541 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     3601 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     3661 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     3721 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     3781 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3841 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3901 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     3961 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4021 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4081 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4141 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     4201 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4261 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4321 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4381 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     4441 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4501 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4561 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     4621 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     4681 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     4741 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta gagcggccgc cccacacacc
     4801 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     4861 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     4921 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     4981 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     5041 tttttttttt gatttttttc tctttcgatg acctcccatt gat
//
