LOCUS       sequence-291576-        5002 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 5002)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5002)
  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..5002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             677..694
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             695..712
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(757..775)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1126..1929
                     /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      2012..2994
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3072..3101
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3123..3710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3722..3749)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3773..3964)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3965..4756,4757..4825))
                     /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     4588..4607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4826..4928)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4929..4960)
                     /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 tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgaaaatgt cagatgtaaa gtgcacatca gtagtcttac tctcagtttt
      481 gcaacaactc agagtagaat catcatctaa attgtgggct caatgtgtcc agttacacaa
      541 tgacattctc ttagctaaag atactactga agcctttgaa aaaatggttt cactactttc
      601 tgttttgctt tccatgcagg gtgctgtaga cataaacaag ctttgtgaag aaatgctgga
      661 caacagggca accttactcg tgccgagagg ttctcaccat catcaccatc attaaacagg
      721 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
      781 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
      841 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
      901 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
      961 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     1021 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     1081 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     1141 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     1201 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     1261 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     1321 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     1381 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     1441 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     1501 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     1561 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     1621 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     1681 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     1741 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     1801 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     1861 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     1921 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     1981 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     2041 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2101 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     2161 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2221 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2281 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2341 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2401 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2461 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2521 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     2581 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     2641 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     2701 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     2761 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     2821 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     2881 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     2941 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     3001 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     3061 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     3121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     3241 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3601 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3661 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3721 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     3781 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3841 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3901 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3961 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     4081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4141 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     4201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4381 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     4441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     4501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4621 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     4681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4801 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     4861 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     4921 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     4981 atgatttaaa tggtcagtta ga
//
