LOCUS       sequence-297593-        6567 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 6567)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6567)
  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-m4046622
FEATURES             Location/Qualifiers
     source          1..6567
                     /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"
     primer_bind     complement(2322..2340)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2691..3494
                     /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      3577..4559
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4637..4666
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4688..5275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5287..5314)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5338..5529)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5530..6321,6322..6390))
                     /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     6153..6172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6391..6493)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6494..6525)
                     /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 gttggggctt gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt
      541 agaccattct tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta
      601 gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact
      661 caactttact taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt
      721 ccattgtgtg ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat
      781 aatgttactg actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt
      841 ttagctaaca cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact
      901 gaggagacat ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga
      961 gaattacatc tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc
     1021 tttactggtt atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa
     1081 aaaggtgact atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt
     1141 ggtgattatt ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg
     1201 ccacaagagc actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag
     1261 ttttctagca atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag
     1321 ggaccacctg gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct
     1381 gctcgcatag tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca
     1441 ttaaaatatt tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag
     1501 tgttttgata aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat
     1561 gcattgcctg agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat
     1621 tatgatttga gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac
     1681 cctgctcaat tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat
     1741 ttcaattcag tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt
     1801 cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt
     1861 aaagcacata aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg
     1921 catgatgttt catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca
     1981 cgtaaccctg cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta
     2041 gcctcaaaga ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat
     2101 gactatgtca tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt
     2161 aatgttgcta ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt
     2221 tatgacaagt tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttataa
     2281 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     2341 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2401 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     2461 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     2521 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     2581 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     2641 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     2701 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     2761 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     2821 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     2881 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     2941 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     3001 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     3061 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     3121 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     3181 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     3241 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     3301 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     3361 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     3421 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     3481 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     3541 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     3601 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3661 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     3721 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     3781 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     3841 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     3901 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     3961 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     4021 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     4081 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     4141 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     4201 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     4261 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     4321 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     4381 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     4441 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     4501 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     4561 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     4621 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     4681 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4741 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     4801 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4861 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4921 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4981 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5041 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5101 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5161 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5221 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5281 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     5341 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5401 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5461 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5521 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5581 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     5641 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5701 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     5761 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5821 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5881 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5941 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     6001 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     6061 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6121 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6181 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     6241 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6301 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6361 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     6421 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     6481 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     6541 caggtatgat ttaaatggtc agttaga
//
