LOCUS       sequence-297849-        6657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6657)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6657)
  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-m4046415
FEATURES             Location/Qualifiers
     source          1..6657
                     /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             2284..2301
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2302..2367
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2412..2430)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2781..3584
                     /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      3667..4649
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4727..4756
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4778..5365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5377..5404)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5428..5619)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5620..6411,6412..6480))
                     /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     6243..6262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6481..6583)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6584..6615)
                     /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 gcagtcggcg cttgtgtatt atgcaactcc cagacgtcgt tgagatgtgg tgcttgtatt
      541 cgtagacctt tcctttgctg taaatgttgc tatgaccacg ttatctcaac ttctcataag
      601 ctagttctga gcgtcaatcc atacgtgtgt aacgcccccg ggtgtgatgt tacagacgtt
      661 acccaattgt acttgggagg tatgtcttac tattgtaagt cccacaagcc acctatatct
      721 tttccattat gcgctaacgg tcaagtcttc ggtttgtaca aaaatacctg tgttggtagt
      781 gataatgtta ccgactttaa cgccattgct acttgtgatt ggactaacgc gggtgattat
      841 atcttggcta acacttgtac cgagaggctt aagttgttcg ccgctgaaac cttaaaggct
      901 acggaagaaa cattcaaatt gagttacggt atcgctactg tcagagaagt tctatctgat
      961 agagaattgc atctctcctg ggaggtgggt aagccaagac caccgttgaa ccggaattac
     1021 gtctttactg gatacagggt tactaagaac tctaaggttc aaattgggga atataccttt
     1081 gaaaagggcg actacggtga cgccgtagtc tatagaggta ccaccactta caaattgaac
     1141 gtcggtgact atttcgtttt aaccagccac acagtgatgc cactgtcagc gccaacatta
     1201 gttccgcaag agcattacgt tagaattact ggtttgtatc caactctaaa tatctctgat
     1261 gaattttcct ccaatgtggc taactaccaa aaagtaggta tgcaaaagta ctctaccttg
     1321 caaggtccac ctggtactgg gaagtcgcac ttcgctatag gtttggcact gtactaccca
     1381 tctgctagaa ttgtttacac ggcatgttct catgctgctg tagacgcctt gtgtgaaaag
     1441 gccttgaaat atttacccat tgataaatgt tcaagaatta tcccagcacg agctcgtgtt
     1501 gaatgtttcg acaagtttaa ggtcaacagc acactagaac aatacgtctt ctgcactgtt
     1561 aatgctttgc cagaaacgac tgctgatatc gtcgtctttg atgaaatctc gatggccact
     1621 aactatgact tatccgtagt taatgctaga cttcgtgcaa agcactacgt ctatattggt
     1681 gatccagctc agctgcctgc accaagaact ttgttaacca agggtacctt ggaaccagaa
     1741 tacttcaact ctgtgtgtcg cctgatgaag acaatcggcc ctgatatgtt cctaggtaca
     1801 tgtagacgtt gtccagctga aattgtggac acggtcagtg ccttggttta cgataacaaa
     1861 ttaaaggcgc ataaggacaa aagcgctcaa tgtttcaaga tgttttacaa gggtgtcatt
     1921 actcacgatg ttagttcggc catcaataga ccccaaatag gtgttgttag agaatttttg
     1981 acaaggaacc cagcatggag aaaggctgtt ttcatatccc catacaattc tcaaaacgcg
     2041 gtagcctcaa aaatcttggg tcttcccact cagaccgttg attcttccca aggaagtgaa
     2101 tatgactacg tgattttcac tcaaaccacc gagactgccc actcttgcaa cgtcaatcgt
     2161 ttcaacgttg ctattacgag agccaaggtt ggcatcttgt gtatcatgtc tgatagagac
     2221 ttgtacgata aattacagtt tacatcctta gaaataccaa gacgaaacgt cgctactttg
     2281 caactcgtgc cgagaggttc tgattataaa gatcatgatg gtgattataa agatcatgat
     2341 attgattata aagatgatga tgataaataa acaggcccct tttcctttgt cgatatcatg
     2401 taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag
     2461 gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag
     2521 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg
     2581 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
     2641 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     2701 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     2761 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     2821 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     2881 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     2941 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     3001 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     3061 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     3121 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     3181 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     3241 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     3301 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     3361 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     3421 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     3481 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     3541 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     3601 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     3661 ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     3721 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     3781 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     3841 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     3901 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     3961 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     4021 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     4081 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     4141 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     4201 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     4261 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     4321 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     4381 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     4441 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     4501 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     4561 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa
     4621 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg
     4681 tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
     4741 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
     4801 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4861 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
     4921 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4981 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5041 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5101 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5161 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5221 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta
     5281 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
     5341 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
     5401 agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc
     5461 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     5521 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5581 gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga
     5641 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5701 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5761 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     5821 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5881 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5941 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6001 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6061 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6121 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     6181 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6241 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6301 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6361 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6421 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6481 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     6541 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     6601 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
