LOCUS       sequence-300433-        6615 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 6615)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6615)
  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-m4046178
FEATURES             Location/Qualifiers
     source          1..6615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      21..1003
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      1081..1110
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1132..1719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1731..1758)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1782..1973)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1974..2765,2766..2834))
                     /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     2597..2616
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2835..2937)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2938..2969)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3036..3477
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3036..3153
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3448..3471
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5295..5312
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5313..5336
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5381..5399)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             5750..6553
                     /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"
ORIGIN
        1 atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
       61 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
      121 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
      181 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
      241 gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta
      301 caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct
      361 cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg
      421 ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt
      481 cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc
      541 cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt
      601 gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact
      661 acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta
      721 ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga
      781 gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac
      841 agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat
      901 tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg
      961 cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt
     1021 cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact
     1081 ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc
     1141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1201 ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg
     1261 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1321 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1381 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1441 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1501 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1561 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1621 gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga
     1681 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg
     1741 cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca
     1801 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     1861 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     1921 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa
     1981 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2041 tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct
     2101 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2161 gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt
     2221 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2281 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2341 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2401 tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt
     2461 atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact
     2521 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2581 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2641 ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg
     2701 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2761 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2821 tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     2881 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt
     2941 caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat
     3001 ggtcagttag agcggccgct tgctatattg aagtacggat tagaagccgc cgagcgggcg
     3061 acagccctcc gacggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg
     3121 aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct
     3181 tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaattaac
     3241 gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg
     3301 taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat ggaaaagctg
     3361 cataaccact ttaactaata ctttcaacat tttcagtttg tattacttct tattcaaatg
     3421 tcataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa
     3481 aaactataat ggcagtcggc gcttgtgtat tatgcaactc ccagacgtcg ttgagatgtg
     3541 gtgcttgtat tcgtagacct ttcctttgct gtaaatgttg ctatgaccac gttatctcaa
     3601 cttctcataa gctagttctg agcgtcaatc catacgtgtg taacgccccc gggtgtgatg
     3661 ttacagacgt tacccaattg tacttgggag gtatgtctta ctattgtaag tcccacaagc
     3721 cacctatatc ttttccatta tgcgctaacg gtcaagtctt cggtttgtac aaaaatacct
     3781 gtgttggtag tgataatgtt accgacttta acgccattgc tacttgtgat tggactaacg
     3841 cgggtgatta tatcttggct aacacttgta ccgagaggct taagttgttc gccgctgaaa
     3901 ccttaaaggc tacggaagaa acattcaaat tgagttacgg tatcgctact gtcagagaag
     3961 ttctatctga tagagaattg catctctcct gggaggtggg taagccaaga ccaccgttga
     4021 accggaatta cgtctttact ggatacaggg ttactaagaa ctctaaggtt caaattgggg
     4081 aatatacctt tgaaaagggc gactacggtg acgccgtagt ctatagaggt accaccactt
     4141 acaaattgaa cgtcggtgac tatttcgttt taaccagcca cacagtgatg ccactgtcag
     4201 cgccaacatt agttccgcaa gagcattacg ttagaattac tggtttgtat ccaactctaa
     4261 atatctctga tgaattttcc tccaatgtgg ctaactacca aaaagtaggt atgcaaaagt
     4321 actctacctt gcaaggtcca cctggtactg ggaagtcgca cttcgctata ggtttggcac
     4381 tgtactaccc atctgctaga attgtttaca cggcatgttc tcatgctgct gtagacgcct
     4441 tgtgtgaaaa ggccttgaaa tatttaccca ttgataaatg ttcaagaatt atcccagcac
     4501 gagctcgtgt tgaatgtttc gacaagttta aggtcaacag cacactagaa caatacgtct
     4561 tctgcactgt taatgctttg ccagaaacga ctgctgatat cgtcgtcttt gatgaaatct
     4621 cgatggccac taactatgac ttatccgtag ttaatgctag acttcgtgca aagcactacg
     4681 tctatattgg tgatccagct cagctgcctg caccaagaac tttgttaacc aagggtacct
     4741 tggaaccaga atacttcaac tctgtgtgtc gcctgatgaa gacaatcggc cctgatatgt
     4801 tcctaggtac atgtagacgt tgtccagctg aaattgtgga cacggtcagt gccttggttt
     4861 acgataacaa attaaaggcg cataaggaca aaagcgctca atgtttcaag atgttttaca
     4921 agggtgtcat tactcacgat gttagttcgg ccatcaatag accccaaata ggtgttgtta
     4981 gagaattttt gacaaggaac ccagcatgga gaaaggctgt tttcatatcc ccatacaatt
     5041 ctcaaaacgc ggtagcctca aaaatcttgg gtcttcccac tcagaccgtt gattcttccc
     5101 aaggaagtga atatgactac gtgattttca ctcaaaccac cgagactgcc cactcttgca
     5161 acgtcaatcg tttcaacgtt gctattacga gagccaaggt tggcatcttg tgtatcatgt
     5221 ctgatagaga cttgtacgat aaattacagt ttacatcctt agaaatacca agacgaaacg
     5281 tcgctacttt gcaactcgtg ccgagaggtt cttggagcca tccgcagttt gaaaaataaa
     5341 caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc
     5401 ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     5461 ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     5521 tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc
     5581 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     5641 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     5701 cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc
     5761 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     5821 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     5881 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     5941 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     6001 caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     6061 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     6121 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     6181 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     6241 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
     6301 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     6361 gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt
     6421 ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     6481 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     6541 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     6601 agcttcaatt taatt
//
