LOCUS       sequence-298143-        4973 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4973)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4973)
  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-m4046223
FEATURES             Location/Qualifiers
     source          1..4973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             648..665
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             666..683
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(728..746)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1097..1900
                     /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      1983..2965
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3043..3072
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3094..3681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3693..3720)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3744..3935)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3936..4727,4728..4796))
                     /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     4559..4578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4797..4899)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4900..4931)
                     /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 tttttcttcc gtttcttttt cttgaaaatt tttttttttg
      241 atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt
      301 ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta
      361 gaaagaaagc atagcaatct aatctaagtt ttaattacaa aatgaaaatg tcagatgtaa
      421 agtgcacatc agtagtctta ctctcagttt tgcaacaact cagagtagaa tcatcatcta
      481 aattgtgggc tcaatgtgtc cagttacaca atgacattct cttagctaaa gatactactg
      541 aagcctttga aaaaatggtt tcactacttt ctgttttgct ttccatgcag ggtgctgtag
      601 acataaacaa gctttgtgaa gaaatgctgg acaacagggc aaccttactc gtgccgagag
      661 gttctcacca tcatcaccat cattaaacag gccccttttc ctttgtcgat atcatgtaat
      721 tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag
      781 gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt
      841 aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc
      901 actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca
      961 gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt
     1021 tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag
     1081 gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag
     1141 tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc
     1201 attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat
     1261 ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa
     1321 gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc
     1381 tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg
     1441 ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca
     1501 ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg
     1561 caagggctcc ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga
     1621 caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta
     1681 cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca
     1741 acagtataga accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag
     1801 aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg
     1861 ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa
     1921 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga
     1981 gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt
     2041 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat
     2101 tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct
     2161 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     2221 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     2281 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     2341 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     2401 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     2461 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     2521 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     2581 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     2641 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     2701 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     2761 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     2821 gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag
     2881 tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc
     2941 gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt
     3001 acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg
     3061 ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3181 gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc
     3721 aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3781 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     3841 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     3901 gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca
     3961 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4021 ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4081 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc
     4141 agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4201 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4261 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4321 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4381 gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4441 tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag
     4501 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4561 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4621 cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4681 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4741 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta
     4801 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     4861 tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga
     4921 ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
