LOCUS       sequence-297727-        7689 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 7689)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7689)
  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-m4046552
FEATURES             Location/Qualifiers
     source          1..7689
                     /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"
     primer_bind     complement(3444..3462)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3813..4616
                     /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      4699..5681
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5759..5788
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5810..6397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6409..6436)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6460..6651)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6652..7443,7444..7512))
                     /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     7275..7294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7513..7615)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7616..7647)
                     /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 taaaatcgag gaaggtaagc tagtcatttg gataaatgga
     2341 gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga cacaggtatc
     2401 aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt tgctgctact
     2461 ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata tgctcagtca
     2521 ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta cccattcact
     2581 tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt tgaagcctta
     2641 tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga agaaattcca
     2701 gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa cttacaagaa
     2761 ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa atacgaaaac
     2821 ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc tggtttgacc
     2881 ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta ctctatcgcc
     2941 gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg ggcctggagt
     3001 aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt caaaggccaa
     3061 ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc tccaaacaag
     3121 gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt ggaggccgtt
     3181 aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga attggctaag
     3241 gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat gccaaacatc
     3301 ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc tgcttccggt
     3361 agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc cttttccttt
     3421 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     3481 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     3541 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     3601 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     3661 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     3721 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     3781 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     3841 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     3901 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     3961 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     4021 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     4081 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     4141 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     4201 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     4261 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     4321 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     4381 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     4441 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     4501 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     4561 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     4621 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     4681 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     4741 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     4801 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     4861 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     4921 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     4981 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     5041 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     5101 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     5161 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     5221 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     5281 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     5341 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     5401 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     5461 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     5521 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     5581 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     5641 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     5701 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     5761 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     5821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5881 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     5941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     6361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     6421 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     6481 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     6541 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     6601 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     6661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6721 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     6781 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6841 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     6901 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6961 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7021 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7081 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     7141 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     7201 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7261 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7321 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7381 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7441 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7501 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     7561 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     7621 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     7681 tcagttaga
//
