LOCUS       sequence-298227-        4878 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4878)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4878)
  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..4878
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             547..564
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             565..588
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(633..651)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1002..1805
                     /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      1888..2870
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2948..2977
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2999..3586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3598..3625)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3649..3840)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3841..4632,4633..4701))
                     /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     4464..4483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4702..4804)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4805..4836)
                     /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 caataaattt ctttttcttg
      241 aaaatttttt tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat
      301 aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta caactttttt
      361 tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttaa ttacaaaatg
      421 attgaacttt cattaattga cttctatttg tgctttttag cctttctgct attccttgtt
      481 ttaattatgc ttattatctt ttggttctca cttgaactgc aagatcataa tgaaacttgt
      541 cacgccctcg tgccgagagg ttcttggagc catccgcagt ttgaaaaata aacaggcccc
      601 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
      661 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
      721 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
      781 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
      841 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
      901 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
      961 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     1021 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     1081 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     1141 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     1201 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     1261 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     1321 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     1381 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     1441 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     1501 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     1561 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     1621 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     1681 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     1741 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     1801 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     1861 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     1921 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     1981 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2041 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2101 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     2161 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     2221 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     2281 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     2341 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     2401 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     2461 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     2521 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     2581 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     2641 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     2701 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     2761 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     2821 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     2881 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     2941 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     3001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3061 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     3121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3481 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3541 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3601 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     3661 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     3721 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     3781 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     3841 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3901 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     3961 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4021 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     4081 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4141 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4201 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4261 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     4321 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     4381 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4441 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4501 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     4561 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4621 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4681 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     4741 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     4801 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     4861 tttaaatggt cagttaga
//
