LOCUS       sequence-291582-        4891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4891)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4891)
  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-m4046356
FEATURES             Location/Qualifiers
     source          1..4891
                     /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             560..577
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             578..601
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(646..664)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1015..1818
                     /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      1901..2883
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2961..2990
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3012..3599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3611..3638)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3662..3853)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3854..4645,4646..4714))
                     /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     4477..4496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4715..4817)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4818..4849)
                     /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 atgattgaac tttcattaat tgacttctat ttgtgctttt tagcctttct
      481 gctattcctt gttttaatta tgcttattat cttttggttc tcacttgaac tgcaagatca
      541 taatgaaact tgtcacgccc tcgtgccgag aggttcttgg agccatccgc agtttgaaaa
      601 ataaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
      661 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
      721 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
      781 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
      841 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
      901 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
      961 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     1021 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     1081 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     1141 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     1201 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     1261 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     1321 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     1381 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     1441 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     1501 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     1561 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     1621 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     1681 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     1741 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     1801 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     1861 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     1921 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     1981 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2041 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2101 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     2161 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     2221 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     2281 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     2341 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     2401 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     2461 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     2521 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     2581 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     2641 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     2701 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     2761 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     2821 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     2881 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     2941 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     3001 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3061 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     3121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3481 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3541 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3601 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     3661 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3721 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3781 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3841 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     3961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4021 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     4081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     4321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4381 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     4561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4681 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     4741 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     4801 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     4861 aggtcaggta tgatttaaat ggtcagttag a
//
