LOCUS       sequence-297804-        5019 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 5019)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5019)
  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-m4046687
FEATURES             Location/Qualifiers
     source          1..5019
                     /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"
     primer_bind     complement(774..792)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1143..1946
                     /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      2029..3011
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3089..3118
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3140..3727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3739..3766)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3790..3981)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3982..4773,4774..4842))
                     /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     4605..4624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4843..4945)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4946..4977)
                     /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 actttacaat aaacaggccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc
      781 ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
      841 gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta
      901 tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta gtgcactgca
      961 gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     1021 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     1081 attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa
     1141 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     1201 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     1261 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     1321 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     1381 tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata
     1441 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     1501 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     1561 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     1621 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     1681 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     1741 tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg
     1801 tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     1861 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     1921 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     1981 actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa cgaagcatct
     2041 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2101 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     2161 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2221 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     2281 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     2341 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     2401 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     2461 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     2521 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     2581 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     2641 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     2701 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     2761 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     2821 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     2881 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     2941 ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     3001 atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa gcttgactcc
     3061 agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt
     3121 agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3181 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3241 ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3601 aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata gtcctgtcgg
     3661 gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3721 atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt
     3781 gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3841 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3901 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3961 ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg
     4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4141 tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc
     4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa
     4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4501 gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag
     4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc
     4861 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     4921 gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc
     4981 tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
