LOCUS       sequence-300483-        6651 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 6651)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6651)
  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-m4046343
FEATURES             Location/Qualifiers
     source          1..6651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      61..1043
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      1121..1150
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1172..1759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1771..1798)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1822..2013)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(2014..2805,2806..2874))
                     /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     2637..2656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2875..2977)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2978..3009)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3076..3517
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3076..3193
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3488..3511
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5329..5346
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5347..5412
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5457..5475)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             5826..6629
                     /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 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
       61 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
      121 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
      181 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
      241 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
      301 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
      361 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
      421 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
      481 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
      541 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
      601 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
      661 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
      721 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
      781 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
      841 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
      901 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
      961 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     1021 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     1081 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     1141 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1201 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1261 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     1321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1621 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     1681 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     1741 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     1801 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     1861 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     1921 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     1981 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     2041 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2101 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2161 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     2221 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2281 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2341 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2401 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2461 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2521 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     2581 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2641 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2701 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2761 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2821 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     2881 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     2941 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     3001 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag agcggccgct
     3061 tgctatattg aagtacggat tagaagccgc cgagcgggcg acagccctcc gacggaagac
     3121 tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg
     3181 ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta tgaagaggaa
     3241 aaattggcag taacctggcc ccacaaacct tcaaattaac gaatcaaatt aacaaccata
     3301 ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat
     3361 gatttttgat ctattaacag atatataaat ggaaaagctg cataaccact ttaactaata
     3421 ctttcaacat tttcagtttg tattacttct tattcaaatg tcataaaagt atcaacaaaa
     3481 aattgttaat atacctctat actttaacgt caaggagaaa aaactataat ggttggggct
     3541 tgtgttcttt gcaattcaca gacttcatta agatgtggtg cttgcatacg tagaccattc
     3601 ttatgttgta aatgctgtta cgaccatgtc atatcaacat cacataaatt agtcttgtct
     3661 gttaatccgt atgtttgcaa tgctccaggt tgtgatgtca cagatgtgac tcaactttac
     3721 ttaggaggta tgagctatta ttgtaaatca cataaaccac ccattagttt tccattgtgt
     3781 gctaatggac aagtttttgg tttatataaa aatacatgtg ttggtagcga taatgttact
     3841 gactttaatg caattgcaac atgtgactgg acaaatgctg gtgattacat tttagctaac
     3901 acctgtactg aaagactcaa gctttttgca gcagaaacgc tcaaagctac tgaggagaca
     3961 tttaaactgt cttatggtat tgctactgta cgtgaagtgc tgtctgacag agaattacat
     4021 ctttcatggg aagttggtaa acctagacca ccacttaacc gaaattatgt ctttactggt
     4081 tatcgtgtaa ctaaaaacag taaagtacaa ataggagagt acacctttga aaaaggtgac
     4141 tatggtgatg ctgttgttta ccgaggtaca acaacttaca aattaaatgt tggtgattat
     4201 tttgtgctga catcacatac agtaatgcca ttaagtgcac ctacactagt gccacaagag
     4261 cactatgtta gaattactgg cttataccca acactcaata tctcagatga gttttctagc
     4321 aatgttgcaa attatcaaaa ggttggtatg caaaagtatt ctacactcca gggaccacct
     4381 ggtactggta agagtcattt tgctattggc ctagctctct actacccttc tgctcgcata
     4441 gtgtatacag cttgctctca tgccgctgtt gatgcactat gtgagaaggc attaaaatat
     4501 ttgcctatag ataaatgtag tagaattata cctgcacgtg ctcgtgtaga gtgttttgat
     4561 aaattcaaag tgaattcaac attagaacag tatgtctttt gtactgtaaa tgcattgcct
     4621 gagacgacag cagatatagt tgtctttgat gaaatttcaa tggccacaaa ttatgatttg
     4681 agtgttgtca atgccagatt acgtgctaag cactatgtgt acattggcga ccctgctcaa
     4741 ttacctgcac cacgcacatt gctaactaag ggcacactag aaccagaata tttcaattca
     4801 gtgtgtagac ttatgaaaac tataggtcca gacatgttcc tcggaacttg tcggcgttgt
     4861 cctgctgaaa ttgttgacac tgtgagtgct ttggtttatg ataataagct taaagcacat
     4921 aaagacaaat cagctcaatg ctttaaaatg ttttataagg gtgttatcac gcatgatgtt
     4981 tcatctgcaa ttaacaggcc acaaataggc gtggtaagag aattccttac acgtaaccct
     5041 gcttggagaa aagctgtctt tatttcacct tataattcac agaatgctgt agcctcaaag
     5101 attttgggac taccaactca aactgttgat tcatcacagg gctcagaata tgactatgtc
     5161 atattcactc aaaccactga aacagctcac tcttgtaatg taaacagatt taatgttgct
     5221 attaccagag caaaagtagg catactttgc ataatgtctg atagagacct ttatgacaag
     5281 ttgcaattta caagtcttga aattccacgt aggaatgtgg caactttact cgtgccgaga
     5341 ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga ttataaagat
     5401 gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     5461 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     5521 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     5581 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     5641 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     5701 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     5761 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     5821 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     5881 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     5941 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     6001 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     6061 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     6121 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     6181 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     6241 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     6301 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     6361 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     6421 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     6481 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     6541 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     6601 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat g
//
