LOCUS       sequence-297655-        5502 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 5502)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5502)
  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-m4046545
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      1..588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(600..627)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(651..842)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(843..1634,1635..1703))
                     /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     1466..1485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1704..1806)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1807..1838)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1905..2346
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1905..2022
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2317..2340
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4158..4175
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4176..4193
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4238..4256)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4607..5410
                     /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
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWXAYLRRCGQQN"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc
      121 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
      181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      481 cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      541 gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      601 gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt
      661 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
      721 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
      781 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
      841 acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
      901 atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc
      961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1021 aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc
     1081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1261 gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca
     1321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt
     1381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1501 ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga
     1561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1681 acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca
     1741 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     1801 cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat
     1861 gatttaaatg gtcagttaga gcggccgctt gctatattga agtacggatt agaagccgcc
     1921 gagcgggcga cagccctccg acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc
     1981 gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca
     2041 atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt
     2101 caaattaacg aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta
     2161 tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg
     2221 gaaaagctgc ataaccactt taactaatac tttcaacatt ttcagtttgt attacttctt
     2281 attcaaatgt cataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc
     2341 aaggagaaaa aactataatg gttggggctt gtgttctttg caattcacag acttcattaa
     2401 gatgtggtgc ttgcatacgt agaccattct tatgttgtaa atgctgttac gaccatgtca
     2461 tatcaacatc acataaatta gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt
     2521 gtgatgtcac agatgtgact caactttact taggaggtat gagctattat tgtaaatcac
     2581 ataaaccacc cattagtttt ccattgtgtg ctaatggaca agtttttggt ttatataaaa
     2641 atacatgtgt tggtagcgat aatgttactg actttaatgc aattgcaaca tgtgactgga
     2701 caaatgctgg tgattacatt ttagctaaca cctgtactga aagactcaag ctttttgcag
     2761 cagaaacgct caaagctact gaggagacat ttaaactgtc ttatggtatt gctactgtac
     2821 gtgaagtgct gtctgacaga gaattacatc tttcatggga agttggtaaa cctagaccac
     2881 cacttaaccg aaattatgtc tttactggtt atcgtgtaac taaaaacagt aaagtacaaa
     2941 taggagagta cacctttgaa aaaggtgact atggtgatgc tgttgtttac cgaggtacaa
     3001 caacttacaa attaaatgtt ggtgattatt ttgtgctgac atcacataca gtaatgccat
     3061 taagtgcacc tacactagtg ccacaagagc actatgttag aattactggc ttatacccaa
     3121 cactcaatat ctcagatgag ttttctagca atgttgcaaa ttatcaaaag gttggtatgc
     3181 aaaagtattc tacactccag ggaccacctg gtactggtaa gagtcatttt gctattggcc
     3241 tagctctcta ctacccttct gctcgcatag tgtatacagc ttgctctcat gccgctgttg
     3301 atgcactatg tgagaaggca ttaaaatatt tgcctataga taaatgtagt agaattatac
     3361 ctgcacgtgc tcgtgtagag tgttttgata aattcaaagt gaattcaaca ttagaacagt
     3421 atgtcttttg tactgtaaat gcattgcctg agacgacagc agatatagtt gtctttgatg
     3481 aaatttcaat ggccacaaat tatgatttga gtgttgtcaa tgccagatta cgtgctaagc
     3541 actatgtgta cattggcgac cctgctcaat tacctgcacc acgcacattg ctaactaagg
     3601 gcacactaga accagaatat ttcaattcag tgtgtagact tatgaaaact ataggtccag
     3661 acatgttcct cggaacttgt cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt
     3721 tggtttatga taataagctt aaagcacata aagacaaatc agctcaatgc tttaaaatgt
     3781 tttataaggg tgttatcacg catgatgttt catctgcaat taacaggcca caaataggcg
     3841 tggtaagaga attccttaca cgtaaccctg cttggagaaa agctgtcttt atttcacctt
     3901 ataattcaca gaatgctgta gcctcaaaga ttttgggact accaactcaa actgttgatt
     3961 catcacaggg ctcagaatat gactatgtca tattcactca aaccactgaa acagctcact
     4021 cttgtaatgt aaacagattt aatgttgcta ttaccagagc aaaagtaggc atactttgca
     4081 taatgtctga tagagacctt tatgacaagt tgcaatttac aagtcttgaa attccacgta
     4141 ggaatgtggc aactttactc gtgccgagag gttctcacca tcatcaccat cattaaacag
     4201 gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc
     4261 ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     4321 tttatttttt ttaatagtta tgtkagtmtt aagaacgtta tttatatttc aaatttttct
     4381 tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg
     4441 aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact
     4501 tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt
     4561 aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac
     4621 atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat
     4681 gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact
     4741 ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt
     4801 gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa
     4861 ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta
     4921 ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt
     4981 gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag
     5041 aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac
     5101 taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca
     5161 aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg
     5221 tttagatgac aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc
     5281 tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa
     5341 ggtagagggt gaacgttaca gaaaagcagg ctgggangca tatttgagaa gatgcggcca
     5401 gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc
     5461 ttcaatttaa ttatatcagt tattacccga gtaacgaagc at
//
