LOCUS       sequence-287678-        5049 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 5049)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5049)
  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-m4046220
FEATURES             Location/Qualifiers
     source          1..5049
                     /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             724..741
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             742..759
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(804..822)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1173..1976
                     /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      2059..3041
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3119..3148
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3170..3757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3769..3796)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3820..4011)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4012..4803,4804..4872))
                     /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     4635..4654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4873..4975)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4976..5007)
                     /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 aaaatgtcag atgtaaagtg cacatcagta gtcttactct cagttttgca acaactcaga
      541 gtagaatcat catctaaatt gtgggctcaa tgtgtccagt tacacaatga cattctctta
      601 gctaaagata ctactgaagc ctttgaaaaa atggtttcac tactttctgt tttgctttcc
      661 atgcagggtg ctgtagacat aaacaagctt tgtgaagaaa tgctggacaa cagggcaacc
      721 ttactcgtgc cgagaggttc tcaccatcat caccatcatt aaacaggccc cttttccttt
      781 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
      841 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
      901 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
      961 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     1021 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1081 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     1141 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     1201 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     1261 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     1321 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     1381 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     1441 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     1501 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     1561 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     1621 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     1681 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     1741 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     1801 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     1861 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     1921 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     1981 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     2041 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     2101 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2161 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     2221 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2281 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     2341 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     2401 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     2461 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     2521 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     2581 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     2641 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     2701 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     2761 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     2821 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     2881 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     2941 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     3001 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     3061 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     3121 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3241 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3301 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3361 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3421 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3481 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3541 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3601 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     3661 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     3721 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     3781 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     3841 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3901 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3961 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     4021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4081 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     4141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4201 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     4261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4441 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     4501 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     4561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4681 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4861 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     4921 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     4981 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     5041 tcagttaga
//
