LOCUS       sequence-298269-        5167 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 5167)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5167)
  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-m4046560
FEATURES             Location/Qualifiers
     source          1..5167
                     /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..877
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(922..940)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1291..2094
                     /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      2177..3159
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3237..3266
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3288..3875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3887..3914)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3938..4129)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4130..4921,4922..4990))
                     /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     4753..4772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4991..5093)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5094..5125)
                     /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 atgaaatttc ttgttttctt aggaatcatc acaactgtag ctgcatttca
      481 ccaagaatgt agtttacagt catgtactca acatcaacca tatgtagttg atgacccgtg
      541 tcctattcac ttctattcta aatggtatat tagagtagga gctagaaaat cagcaccttt
      601 aattgaattg tgcgtggatg aggctggttc taaatcaccc attcagtaca tcgatatcgg
      661 taattataca gtttcctgtt taccttttac aattaattgc caggaaccta aattgggtag
      721 tcttgtagtg cgttgttcgt tctatgaaga ctttttagag tatcatgacg ttcgtgttgt
      781 tttagatttc atcctcgtgc cgagaggttc tgattataaa gatcatgatg gtgattataa
      841 agatcatgat attgattata aagatgatga tgataaataa acaggcccct tttcctttgt
      901 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
      961 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     1021 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     1081 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     1141 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1201 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1261 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1321 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1381 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     1441 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     1501 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     1561 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     1621 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     1681 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     1741 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     1801 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     1861 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     1921 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     1981 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     2041 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     2101 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     2161 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2221 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2281 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2341 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2401 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2461 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     2521 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     2581 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     2641 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     2701 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     2761 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     2821 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     2881 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     2941 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     3001 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     3061 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     3121 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     3181 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     3241 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     3301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3361 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     3421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     3781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     3841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     3901 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     3961 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4021 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4081 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     4141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4201 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     4261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4321 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     4381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4561 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4621 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     4681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4801 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4981 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     5041 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     5101 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     5161 agttaga
//
