LOCUS       sequence-292513-        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-m4046525
FEATURES             Location/Qualifiers
     source          1..5055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        64..505
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    64..181
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     476..499
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             763..780
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             781..804
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(849..867)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1218..2021
                     /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      2104..3086
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3164..3193
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3215..3802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3814..3841)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3865..4056)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4057..4848,4849..4917))
                     /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     4680..4699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4918..5020)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5021..5052)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tgctatggag gtcaggtatg atttaaatgg tcagttagag cggccgcttg ctatattgaa
       61 gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc tcctccgtgc
      121 gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc
      181 gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa attggcagta
      241 acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg atgataatgc
      301 gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga tttttgatct
      361 attaacagat atataaatgg aaaagctgca taaccacttt aactaatact ttcaacattt
      421 tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa ttgttaatat
      481 acctctatac tttaacgtca aggagaaaaa actataatga aaatgtcaga tgtaaagtgc
      541 acatcagtag tcttactctc agttttgcaa caactcagag tagaatcatc atctaaattg
      601 tgggctcaat gtgtccagtt acacaatgac attctcttag ctaaagatac tactgaagcc
      661 tttgaaaaaa tggtttcact actttctgtt ttgctttcca tgcagggtgc tgtagacata
      721 aacaagcttt gtgaagaaat gctggacaac agggcaacct tactcgtgcc gagaggttct
      781 tggagccatc cgcagtttga aaaataaaca ggcccctttt cctttgtcga tatcatgtaa
      841 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
      901 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
      961 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     1021 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     1081 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     1141 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     1201 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     1261 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     1321 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     1381 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     1441 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     1501 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     1561 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     1621 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     1681 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     1741 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     1801 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     1861 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     1921 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     1981 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     2041 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     2101 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     2161 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     2221 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     2281 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     2341 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     2401 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     2461 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     2521 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     2581 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     2641 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     2701 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     2761 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     2821 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     2881 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     2941 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     3001 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     3061 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     3121 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     3181 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3241 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3301 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     3721 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     3781 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     3841 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     3901 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3961 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4021 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     4081 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4141 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4201 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     4261 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4321 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4381 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4441 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4501 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4561 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     4621 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4681 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4741 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4801 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4861 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     4921 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     4981 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     5041 aggcttttga ctttc
//
