LOCUS       sequence-287610-        5014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5014)
  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-m4046166
FEATURES             Location/Qualifiers
     source          1..5014
                     /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             683..700
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             701..724
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(769..787)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1138..1941
                     /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      2024..3006
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3084..3113
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3135..3722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3734..3761)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3785..3976)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3977..4768,4769..4837))
                     /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     4600..4619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4838..4940)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4941..4972)
                     /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 atgtccaaaa tgtcagacgt caagtgtacg tcggtagttc tcctgagtgt
      481 tctacaacag ttacgtgtgg agagctcttc caagctttgg gcacaatgcg tccaattgca
      541 caatgatatt ttattggcca aagatacaac cgaagctttc gaaaagatgg tttctttgtt
      601 gtctgtcttg ttgtctatgc aaggtgctgt tgacataaac aagttgtgtg aagaaatgct
      661 agataacaga gctactttac aactcgtgcc gagaggttct tggagccatc cgcagtttga
      721 aaaataaaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
      781 ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
      841 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
      901 caaatttttc ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca
      961 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     1021 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     1081 ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg
     1141 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     1201 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     1261 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     1321 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     1381 gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc
     1441 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     1501 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     1561 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     1621 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     1681 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     1741 cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat
     1801 gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     1861 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     1921 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     1981 caaattagag cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct
     2041 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     2101 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     2161 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2221 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     2281 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     2341 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     2401 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     2461 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     2521 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     2581 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     2641 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     2701 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     2761 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     2821 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     2881 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     2941 tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     3001 ttctagctag agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt
     3061 aactggactg caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa
     3121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3241 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3601 agcgcacgag ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc
     3661 gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3721 aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc
     3781 ctcatcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3841 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3901 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3961 tttcctgttt ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4021 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg
     4081 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4141 atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt
     4201 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4261 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4321 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4381 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg
     4441 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4501 ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4561 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4621 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct
     4681 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4741 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4801 agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa
     4861 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     4921 aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta
     4981 tggaggtcag gtatgattta aatggtcagt taga
//
