LOCUS       sequence-297878-        6609 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 6609)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6609)
  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-m4046372
FEATURES             Location/Qualifiers
     source          1..6609
                     /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..2319
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2364..2382)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2733..3536
                     /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      3619..4601
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4679..4708
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4730..5317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5329..5356)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5380..5571)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5572..6363,6364..6432))
                     /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     6195..6214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6433..6535)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6536..6567)
                     /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 tcaccatcat caccatcatt aaacaggccc cttttccttt
     2341 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     2401 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     2461 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2521 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     2581 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     2641 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2701 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2761 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     2821 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2881 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2941 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     3001 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     3061 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     3121 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     3181 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     3241 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     3301 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     3361 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     3421 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     3481 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     3541 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3601 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     3661 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3721 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     3781 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     3841 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     3901 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     3961 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     4021 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     4081 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     4141 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     4201 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     4261 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     4321 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     4381 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     4441 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     4501 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     4561 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     4621 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     4681 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     4861 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5101 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     5341 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     5401 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5461 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5521 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5641 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5761 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6001 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     6061 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6241 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6421 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     6481 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     6541 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     6601 tcagttaga
//
