LOCUS       sequence-297217-        6548 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6548)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6548)
  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-m4046435
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             2223..2240
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2241..2258
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2303..2321)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2672..3475
                     /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      3558..4540
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4618..4647
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4669..5256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5268..5295)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5319..5510)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5511..6302,6303..6371))
                     /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     6134..6153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6372..6474)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6475..6506)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 ttttgaaaaw tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag
      301 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
      361 ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca
      421 aaatggttgg ggcttgtgtt ctttgcaatt cacagacttc attaagatgt ggtgcttgca
      481 tacgtagacc attcttatgt tgtaaatgct gttacgacca tgtcatatca acatcacata
      541 aattagtctt gtctgttaat ccgtatgttt gcaatgctcc aggttgtgat gtcacagatg
      601 tgactcaact ttacttagga ggtatgagct attattgtaa atcacataaa ccacccatta
      661 gttttccatt gtgtgctaat ggacaagttt ttggtttata taaaaataca tgtgttggta
      721 gcgataatgt tactgacttt aatgcaattg caacatgtga ctggacaaat gctggtgatt
      781 acattttagc taacacctgt actgaaagac tcaagctttt tgcagcagaa acgctcaaag
      841 ctactgagga gacatttaaa ctgtcttatg gtattgctac tgtacgtgaa gtgctgtctg
      901 acagagaatt acatctttca tgggaagttg gtaaacctag accaccactt aaccgaaatt
      961 atgtctttac tggttatcgt gtaactaaaa acagtaaagt acaaatagga gagtacacct
     1021 ttgaaaaagg tgactatggt gatgctgttg tttaccgagg tacaacaact tacaaattaa
     1081 atgttggtga ttattttgtg ctgacatcac atacagtaat gccattaagt gcacctacac
     1141 tagtgccaca agagcactat gttagaatta ctggcttata cccaacactc aatatctcag
     1201 atgagttttc tagcaatgtt gcaaattatc aaaaggttgg tatgcaaaag tattctacac
     1261 tccagggacc acctggtact ggtaagagtc attttgctat tggcctagct ctctactacc
     1321 cttctgctcg catagtgtat acagcttgct ctcatgccgc tgttgatgca ctatgtgaga
     1381 aggcattaaa atatttgcct atagataaat gtagtagaat tatacctgca cgtgctcgtg
     1441 tagagtgttt tgataaattc aaagtgaatt caacattaga acagtatgtc ttttgtactg
     1501 taaatgcatt gcctgagacg acagcagata tagttgtctt tgatgaaatt tcaatggcca
     1561 caaattatga tttgagtgtt gtcaatgcca gattacgtgc taagcactat gtgtacattg
     1621 gcgaccctgc tcaattacct gcaccacgca cattgctaac taagggcaca ctagaaccag
     1681 aatatttcaa ttcagtgtgt agacttatga aaactatagg tccagacatg ttcctcggaa
     1741 cttgtcggcg ttgtcctgct gaaattgttg acactgtgag tgctttggtt tatgataata
     1801 agcttaaagc acataaagac aaatcagctc aatgctttaa aatgttttat aagggtgtta
     1861 tcacgcatga tgtttcatct gcaattaaca ggccacaaat aggcgtggta agagaattcc
     1921 ttacacgtaa ccctgcttgg agaaaagctg tctttatttc accttataat tcacagaatg
     1981 ctgtagcctc aaagattttg ggactaccaa ctcaaactgt tgattcatca cagggctcag
     2041 aatatgacta tgtcatattc actcaaacca ctgaaacagc tcactcttgt aatgtaaaca
     2101 gatttaatgt tgctattacc agagcaaaag taggcatact ttgcataatg tctgatagag
     2161 acctttatga caagttgcaa tttacaagtc ttgaaattcc acgtaggaat gtggcaactt
     2221 tactcgtgcc gagaggttct caccatcatc accatcatta aacaggcccc ttttcctttg
     2281 tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc
     2341 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat
     2401 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa
     2461 agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc
     2521 ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat
     2581 acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag
     2641 aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc
     2701 tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac
     2761 aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc
     2821 attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat
     2881 ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga
     2941 ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata
     3001 cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt
     3061 tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt
     3121 agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga
     3181 cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg
     3241 aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg
     3301 agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat
     3361 tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
     3421 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
     3481 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
     3541 tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     3601 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3661 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     3721 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3781 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     3841 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     3901 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     3961 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     4021 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     4081 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     4141 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     4201 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     4261 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     4321 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     4381 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     4441 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt
     4501 ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc
     4561 gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc
     4621 tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt
     4681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4741 tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag
     4801 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta
     5161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt
     5221 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc
     5281 acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca
     5341 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     5401 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     5461 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc
     5521 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5581 ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca
     5641 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5701 gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat
     5761 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5821 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5881 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5941 ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg
     6001 gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt
     6061 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6121 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6181 aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6241 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6301 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6361 tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     6421 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa
     6481 aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt
     6541 cagttaga
//
