LOCUS       sequence-291596-        6199 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 6199)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6199)
  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-m4046243
FEATURES             Location/Qualifiers
     source          1..6199
                     /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"
     primer_bind     complement(1954..1972)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2323..3126
                     /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      3209..4191
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4269..4298
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4320..4907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4919..4946)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4970..5161)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5162..5953,5954..6022))
                     /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     5785..5804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6023..6125)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6126..6157)
                     /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 taaaatcgag gaaggtaagc tagtcatttg
      841 gataaatgga gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga
      901 cacaggtatc aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt
      961 tgctgctact ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata
     1021 tgctcagtca ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta
     1081 cccattcact tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt
     1141 tgaagcctta tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga
     1201 agaaattcca gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa
     1261 cttacaagaa ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa
     1321 atacgaaaac ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc
     1381 tggtttgacc ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta
     1441 ctctatcgcc gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg
     1501 ggcctggagt aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt
     1561 caaaggccaa ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc
     1621 tccaaacaag gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt
     1681 ggaggccgtt aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga
     1741 attggctaag gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat
     1801 gccaaacatc ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc
     1861 tgcttccggt agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc
     1921 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     1981 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     2041 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     2101 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     2161 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     2221 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     2281 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     2341 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     2401 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     2461 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     2521 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     2581 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     2641 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     2701 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     2761 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     2821 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     2881 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     2941 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     3001 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     3061 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     3121 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     3181 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     3241 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     3301 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     3361 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3421 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     3481 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     3541 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     3601 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     3661 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     3721 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     3781 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     3841 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     3901 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     3961 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     4021 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     4081 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     4141 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     4201 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     4261 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     4321 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4381 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     4441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4681 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4741 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4801 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4861 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4921 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     4981 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     5041 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     5101 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     5161 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5221 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     5281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5341 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     5401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5581 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     5641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     5701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5821 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     5881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6001 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     6061 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     6121 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     6181 atttaaatgg tcagttaga
//
