LOCUS       sequence-299940-        5119 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 5119)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5119)
  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-m4046316
FEATURES             Location/Qualifiers
     source          1..5119
                     /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..829
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(874..892)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1243..2046
                     /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      2129..3111
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3189..3218
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3240..3827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3839..3866)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3890..4081)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4082..4873,4874..4942))
                     /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     4705..4724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4943..5045)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5046..5077)
                     /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 tcaccatcat caccatcatt aaacaggccc
      841 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
      901 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
      961 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1021 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     1081 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     1141 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     1201 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     1261 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     1321 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     1381 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     1441 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     1501 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     1561 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     1621 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     1681 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     1741 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     1801 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     1861 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     1921 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     1981 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     2041 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     2101 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     2161 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2221 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     2281 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2341 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     2401 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     2461 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     2521 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     2581 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     2641 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     2701 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     2761 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     2821 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     2881 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     2941 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     3001 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     3061 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     3121 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     3181 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     3241 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3301 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     3361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3721 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3781 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3841 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     3901 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3961 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4021 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4081 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4141 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     4201 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4261 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     4321 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4381 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4441 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4501 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     4561 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     4621 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4681 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4741 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     4801 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4861 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4921 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     4981 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5041 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     5101 atttaaatgg tcagttaga
//
