LOCUS       sequence-298961-        5055 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 5055)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5055)
  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-m4046165
FEATURES             Location/Qualifiers
     source          1..5055
                     /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             730..747
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             748..765
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(810..828)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1179..1982
                     /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      2065..3047
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3125..3154
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3176..3763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3775..3802)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3826..4017)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4018..4809,4810..4878))
                     /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     4641..4660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4879..4981)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4982..5013)
                     /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 tccaaaatgt cagacgtcaa gtgtacgtcg gtagttctcc tgagtgttct acaacagtta
      541 cgtgtggaga gctcttccaa gctttgggca caatgcgtcc aattgcacaa tgatatttta
      601 ttggccaaag atacaaccga agctttcgaa aagatggttt ctttgttgtc tgtcttgttg
      661 tctatgcaag gtgctgttga cataaacaag ttgtgtgaag aaatgctaga taacagagct
      721 actttacaac tcgtgccgag aggttctcac catcatcacc atcattaaac aggccccttt
      781 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tcctcccaca
      841 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
      901 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
      961 tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     1021 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     1081 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     1141 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     1201 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     1261 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     1321 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     1381 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     1441 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     1501 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     1561 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     1621 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     1681 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     1741 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     1801 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     1861 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     1921 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     1981 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     2041 aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     2101 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2161 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     2221 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     2281 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     2341 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     2401 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     2461 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     2521 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     2581 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     2641 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     2701 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     2761 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     2821 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     2881 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     2941 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag
     3001 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga
     3061 acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact
     3121 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag
     3181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag
     3301 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3361 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3421 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3481 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3541 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3601 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3661 aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca
     3721 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga
     3781 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc
     3841 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     3901 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     3961 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta
     4021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4081 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag
     4141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4201 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc
     4261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4441 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat
     4501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt
     4561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4681 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag
     4741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4861 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     4921 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     4981 aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt
     5041 aaatggtcag ttaga
//
