LOCUS       sequence-298240-        4938 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 4938)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4938)
  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-m4046491
FEATURES             Location/Qualifiers
     source          1..4938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             607..624
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             625..648
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(693..711)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1062..1865
                     /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      1948..2930
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3008..3037
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3059..3646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3658..3685)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3709..3900)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3901..4692,4693..4761))
                     /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     4524..4543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4762..4864)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4865..4896)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 attgaacttt cattaattga cttctatttg tgctttttag cctttctgct attccttgtt
      541 ttaattatgc ttattatctt ttggttctca cttgaactgc aagatcataa tgaaacttgt
      601 cacgccctcg tgccgagagg ttcttggagc catccgcagt ttgaaaaata aacaggcccc
      661 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
      721 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
      781 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
      841 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
      901 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
      961 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     1021 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     1081 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     1141 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     1201 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     1261 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     1321 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     1381 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     1441 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     1501 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     1561 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     1621 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     1681 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     1741 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     1801 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     1861 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     1921 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     1981 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2041 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2101 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2161 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     2221 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     2281 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     2341 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     2401 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     2461 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     2521 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     2581 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     2641 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     2701 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     2761 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     2821 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     2881 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     2941 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     3001 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     3061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3121 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     3181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3241 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3541 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3601 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3661 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     3721 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     3781 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     3841 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     3901 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3961 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     4021 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4081 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     4141 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4201 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4261 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4321 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     4381 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     4441 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4501 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4561 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     4621 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4681 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4741 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     4801 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     4861 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     4921 tttaaatggt cagttaga
//
