LOCUS       sequence-298233-        5125 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 5125)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5125)
  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-m4046543
FEATURES             Location/Qualifiers
     source          1..5125
                     /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             794..811
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             812..835
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(880..898)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1249..2052
                     /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      2135..3117
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3195..3224
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3246..3833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3845..3872)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3896..4087)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4088..4879,4880..4948))
                     /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     4711..4730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4949..5051)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5052..5083)
                     /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 atgaaattct tggtttttct tggtatcata actacagtag ccgctttcca
      481 ccaggaatgt tccttacaaa gctgcacgca acatcaaccg tacgtcgttg atgacccatg
      541 tccaatccac ttctattcga agtggtacat tagagttgga gctaggaagt ctgcaccttt
      601 gattgagctg tgtgtcgatg aagctggttc aaagtctcca attcaataca tcgacattgg
      661 taattatact gtctcctgtt tgccctttac cattaactgc caagaaccaa aattaggcag
      721 tttggttgtc cgatgttctt tctacgaaga ctttttggaa tatcatgatg tgcgtgttgt
      781 cctagatttc atcctcgtgc cgagaggttc ttggagccat ccgcagtttg aaaaataaac
      841 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
      901 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
      961 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1021 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
     1081 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     1141 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     1201 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     1261 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     1321 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     1381 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     1441 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     1501 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     1561 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     1621 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     1681 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     1741 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     1801 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     1861 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     1921 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     1981 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     2041 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     2101 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     2161 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     2221 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     2281 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2341 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     2401 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     2461 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     2521 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     2581 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     2641 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     2701 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     2761 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     2821 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     2881 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     2941 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3001 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3061 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     3121 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     3181 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     3241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     3361 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3421 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3481 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3541 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3601 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3661 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3721 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3781 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3841 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     3901 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3961 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4021 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4081 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4141 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     4201 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4261 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     4321 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4381 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4441 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4501 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     4561 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     4621 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4681 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4741 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     4801 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4861 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4921 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     4981 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5041 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     5101 ggtatgattt aaatggtcag ttaga
//
