LOCUS       sequence-292486-        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-m4046416
FEATURES             Location/Qualifiers
     source          1..5125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..415
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             792..809
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             810..833
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(878..896)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1247..2050
                     /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      2133..3115
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3193..3222
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3244..3831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3843..3870)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3894..4085)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4086..4877,4878..4946))
                     /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     4709..4728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4947..5049)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5050..5081)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggccgcccca cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt
       61 tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt
      121 cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa
      181 aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt
      241 tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata
      301 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
      361 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta
      421 attacaaaat gaaatttctt gttttcttag gaatcatcac aactgtagct gcatttcacc
      481 aagaatgtag tttacagtca tgtactcaac atcaaccata tgtagttgat gacccgtgtc
      541 ctattcactt ctattctaaa tggtatatta gagtaggagc tagaaaatca gcacctttaa
      601 ttgaattgtg cgtggatgag gctggttcta aatcacccat tcagtacatc gatatcggta
      661 attatacagt ttcctgttta ccttttacaa ttaattgcca ggaacctaaa ttgggtagtc
      721 ttgtagtgcg ttgttcgttc tatgaagact ttttagagta tcatgacgtt cgtgttgttt
      781 tagatttcat cctcgtgccg agaggttctt ggagccatcc gcagtttgaa aaataaacag
      841 gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc
      901 ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
      961 tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     1021 tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg
     1081 aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact
     1141 tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt
     1201 aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac
     1261 atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat
     1321 gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact
     1381 ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt
     1441 gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa
     1501 ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta
     1561 ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt
     1621 gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag
     1681 aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac
     1741 taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca
     1801 aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg
     1861 tttagatgac aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc
     1921 tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa
     1981 ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca
     2041 gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc
     2101 ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag
     2161 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2221 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     2281 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     2341 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     2401 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     2461 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     2521 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     2581 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     2641 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     2701 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     2761 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     2821 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     2881 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     2941 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     3001 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     3061 gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga
     3121 gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc
     3181 aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc
     3241 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3301 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc
     3361 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3601 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3721 gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3781 cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3841 aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg
     3901 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3961 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     4021 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     4081 gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4141 tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct
     4201 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4261 ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc
     4321 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4381 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4441 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4501 aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta
     4561 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc
     4621 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4681 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4741 gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg
     4801 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4861 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4921 gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac
     4981 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     5041 tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg
     5101 tatgatttaa atggtcagtt agagc
//
