LOCUS       sequence-297714-        6135 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 6135)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6135)
  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-m4046202
FEATURES             Location/Qualifiers
     source          1..6135
                     /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"
     primer_bind     complement(1890..1908)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2259..3062
                     /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      3145..4127
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4205..4234
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4256..4843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4855..4882)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4906..5097)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5098..5889,5890..5958))
                     /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     5721..5740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5959..6061)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6062..6093)
                     /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 aggttctaaa atcgaggaag gtaagctagt catttggata
      781 aatggagata agggttacaa cggtttggca gaagtaggca agaaatttga aaaggacaca
      841 ggtatcaagg tgacggttga acatccagac aagttagaag aaaaattccc tcaagttgct
      901 gctactggtg atggtcccga cattattttc tgggcccacg atcgttttgg tggatatgct
      961 cagtcaggtt tgctggctga gatcacccca gacaaggcct tccaagataa attgtaccca
     1021 ttcacttggg atgctgtcag atacaacggt aagcttatag catatccaat tgctgttgaa
     1081 gccttatcgt tgatctacaa taaggacttg ttacctaacc caccaaagac atgggaagaa
     1141 attccagctt tggataaaga gctaaaggcc aagggaaaaa gcgctttgat gtttaactta
     1201 caagaacctt atttcacctg gccattaatc gctgctgacg gtggttacgc atttaaatac
     1261 gaaaacggta agtatgacat taaggatgtc ggcgttgata atgctggtgc aaaggctggt
     1321 ttgaccttct tggttgattt aataaagaat aaacacatga acgctgacac tgactactct
     1381 atcgccgaag ccgctttcaa caagggtgaa actgctatga ccataaatgg tccatgggcc
     1441 tggagtaaca ttgatacttc taaagtaaac tatggtgtca ctgttttgcc tacattcaaa
     1501 ggccaaccat ccaagccctt tgtcggtgtt ctatccgctg gcatcaatgc tgcttctcca
     1561 aacaaggagc tagctaaaga atttttggaa aattacttgt taactgacga aggtttggag
     1621 gccgttaaca aggacaagcc attgggtgct gtcgctttga aatcttacga agaagaattg
     1681 gctaaggatc ccagaattgc cgctaccatg gagaacgccc aaaagggtga aattatgcca
     1741 aacatcccac aaatgtcagc cttctggtac gcagttagga ctgccgtgat taatgctgct
     1801 tccggtagac aaaccgtcga cgaagcatta aaagatgctc aaacgtaaac aggccccttt
     1861 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tcctcccaca
     1921 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     1981 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     2041 tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     2101 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     2161 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     2221 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     2281 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     2341 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     2401 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     2461 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     2521 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     2581 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     2641 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     2701 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     2761 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     2821 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     2881 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     2941 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     3001 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     3061 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     3121 aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     3181 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     3241 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     3301 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     3361 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     3421 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     3481 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     3541 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     3601 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     3661 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     3721 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     3781 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     3841 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     3901 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     3961 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     4021 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag
     4081 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga
     4141 acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact
     4201 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag
     4261 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4321 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag
     4381 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4441 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4501 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4561 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4621 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4681 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4741 aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca
     4801 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga
     4861 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc
     4921 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     4981 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     5041 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta
     5101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5161 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag
     5221 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5281 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc
     5341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5521 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat
     5581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt
     5641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5761 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag
     5821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5941 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     6001 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     6061 aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt
     6121 aaatggtcag ttaga
//
