LOCUS       sequence-298148-        6592 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6592)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6592)
  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-m4046250
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             2219..2236
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2237..2302
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2347..2365)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2716..3519
                     /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      3602..4584
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4662..4691
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4713..5300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5312..5339)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5363..5554)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5555..6346,6347..6415))
                     /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     6178..6197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6416..6518)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6519..6550)
                     /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 tcgaaaatca cgtttctttt tcttgaaaat
      241 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
      301 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
      361 cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca aaatggcagt
      421 cggcgcttgt gtattatgca actcccagac gtcgttgaga tgtggtgctt gtattcgtag
      481 acctttcctt tgctgtaaat gttgctatga ccacgttatc tcaacttctc ataagctagt
      541 tctgagcgtc aatccatacg tgtgtaacgc ccccgggtgt gatgttacag acgttaccca
      601 attgtacttg ggaggtatgt cttactattg taagtcccac aagccaccta tatcttttcc
      661 attatgcgct aacggtcaag tcttcggttt gtacaaaaat acctgtgttg gtagtgataa
      721 tgttaccgac tttaacgcca ttgctacttg tgattggact aacgcgggtg attatatctt
      781 ggctaacact tgtaccgaga ggcttaagtt gttcgccgct gaaaccttaa aggctacgga
      841 agaaacattc aaattgagtt acggtatcgc tactgtcaga gaagttctat ctgatagaga
      901 attgcatctc tcctgggagg tgggtaagcc aagaccaccg ttgaaccgga attacgtctt
      961 tactggatac agggttacta agaactctaa ggttcaaatt ggggaatata cctttgaaaa
     1021 gggcgactac ggtgacgccg tagtctatag aggtaccacc acttacaaat tgaacgtcgg
     1081 tgactatttc gttttaacca gccacacagt gatgccactg tcagcgccaa cattagttcc
     1141 gcaagagcat tacgttagaa ttactggttt gtatccaact ctaaatatct ctgatgaatt
     1201 ttcctccaat gtggctaact accaaaaagt aggtatgcaa aagtactcta ccttgcaagg
     1261 tccacctggt actgggaagt cgcacttcgc tataggtttg gcactgtact acccatctgc
     1321 tagaattgtt tacacggcat gttctcatgc tgctgtagac gccttgtgtg aaaaggcctt
     1381 gaaatattta cccattgata aatgttcaag aattatccca gcacgagctc gtgttgaatg
     1441 tttcgacaag tttaaggtca acagcacact agaacaatac gtcttctgca ctgttaatgc
     1501 tttgccagaa acgactgctg atatcgtcgt ctttgatgaa atctcgatgg ccactaacta
     1561 tgacttatcc gtagttaatg ctagacttcg tgcaaagcac tacgtctata ttggtgatcc
     1621 agctcagctg cctgcaccaa gaactttgtt aaccaagggt accttggaac cagaatactt
     1681 caactctgtg tgtcgcctga tgaagacaat cggccctgat atgttcctag gtacatgtag
     1741 acgttgtcca gctgaaattg tggacacggt cagtgccttg gtttacgata acaaattaaa
     1801 ggcgcataag gacaaaagcg ctcaatgttt caagatgttt tacaagggtg tcattactca
     1861 cgatgttagt tcggccatca atagacccca aataggtgtt gttagagaat ttttgacaag
     1921 gaacccagca tggagaaagg ctgttttcat atccccatac aattctcaaa acgcggtagc
     1981 ctcaaaaatc ttgggtcttc ccactcagac cgttgattct tcccaaggaa gtgaatatga
     2041 ctacgtgatt ttcactcaaa ccaccgagac tgcccactct tgcaacgtca atcgtttcaa
     2101 cgttgctatt acgagagcca aggttggcat cttgtgtatc atgtctgata gagacttgta
     2161 cgataaatta cagtttacat ccttagaaat accaagacga aacgtcgcta ctttgcaact
     2221 cgtgccgaga ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga
     2281 ttataaagat gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt
     2341 agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg
     2401 agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta
     2461 agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca
     2521 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     2581 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     2641 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     2701 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     2761 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     2821 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     2881 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     2941 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
     3001 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     3061 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     3121 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     3181 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     3241 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     3301 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     3361 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     3421 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     3481 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     3541 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     3601 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     3661 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     3721 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     3781 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     3841 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     3901 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     3961 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     4021 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     4081 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     4141 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     4201 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     4261 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     4321 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     4381 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     4441 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     4501 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
     4561 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
     4621 caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg
     4681 gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4741 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4801 ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc
     4861 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4921 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4981 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5041 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5101 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5161 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt
     5221 atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag
     5281 gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca
     5341 ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     5401 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     5461 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     5521 ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac
     5581 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5641 taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5701 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca
     5761 gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5821 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5881 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5941 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6001 ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6061 ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt
     6121 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6181 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6241 gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6301 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6361 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag
     6421 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     6481 cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag
     6541 gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
