LOCUS       sequence-287693-        5061 bp    DNA     circular SYN 15-APR-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 5061)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5061)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046222
FEATURES             Location/Qualifiers
     source          1..5061
                     /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..771
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(816..834)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1185..1988
                     /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      2071..3053
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3131..3160
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3182..3769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3781..3808)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3832..4023)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4024..4815,4816..4884))
                     /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     4647..4666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4885..4987)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4988..5019)
                     /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 aggttcttgg agccatccgc agtttgaaaa ataaacaggc
      781 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
      841 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
      901 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
      961 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
     1021 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     1081 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     1141 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     1201 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     1261 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     1321 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     1381 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     1441 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     1501 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     1561 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     1621 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     1681 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     1741 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     1801 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     1861 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     1921 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     1981 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     2041 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     2101 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2161 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     2221 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     2281 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     2341 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     2401 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     2461 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     2521 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     2581 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     2641 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     2701 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     2761 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     2821 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     2881 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     2941 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     3001 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     3061 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     3121 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     3181 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3241 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     3301 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3361 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3421 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3481 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3541 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3601 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3661 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3721 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3781 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     3841 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     3901 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     3961 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4021 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4081 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     4141 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4201 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     4261 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4321 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4381 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4441 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     4501 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     4561 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4621 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4681 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     4741 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4801 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4861 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     4921 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     4981 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     5041 tgatttaaat ggtcagttag a
//
