LOCUS       sequence-291657-        6526 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 6526)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6526)
  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-m4046656
FEATURES             Location/Qualifiers
     source          1..6526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2281..2299)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2650..3453
                     /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      3536..4518
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4596..4625
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4647..5234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5246..5273)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5297..5488)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5489..6280,6281..6349))
                     /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     6112..6131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6350..6452)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6453..6484)
                     /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 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggcagtcg gcgcttgtgt attatgcaac tcccagacgt cgttgagatg
      481 tggtgcttgt attcgtagac ctttcctttg ctgtaaatgt tgctatgacc acgttatctc
      541 aacttctcat aagctagttc tgagcgtcaa tccatacgtg tgtaacgccc ccgggtgtga
      601 tgttacagac gttacccaat tgtacttggg aggtatgtct tactattgta agtcccacaa
      661 gccacctata tcttttccat tatgcgctaa cggtcaagtc ttcggtttgt acaaaaatac
      721 ctgtgttggt agtgataatg ttaccgactt taacgccatt gctacttgtg attggactaa
      781 cgcgggtgat tatatcttgg ctaacacttg taccgagagg cttaagttgt tcgccgctga
      841 aaccttaaag gctacggaag aaacattcaa attgagttac ggtatcgcta ctgtcagaga
      901 agttctatct gatagagaat tgcatctctc ctgggaggtg ggtaagccaa gaccaccgtt
      961 gaaccggaat tacgtcttta ctggatacag ggttactaag aactctaagg ttcaaattgg
     1021 ggaatatacc tttgaaaagg gcgactacgg tgacgccgta gtctatagag gtaccaccac
     1081 ttacaaattg aacgtcggtg actatttcgt tttaaccagc cacacagtga tgccactgtc
     1141 agcgccaaca ttagttccgc aagagcatta cgttagaatt actggtttgt atccaactct
     1201 aaatatctct gatgaatttt cctccaatgt ggctaactac caaaaagtag gtatgcaaaa
     1261 gtactctacc ttgcaaggtc cacctggtac tgggaagtcg cacttcgcta taggtttggc
     1321 actgtactac ccatctgcta gaattgttta cacggcatgt tctcatgctg ctgtagacgc
     1381 cttgtgtgaa aaggccttga aatatttacc cattgataaa tgttcaagaa ttatcccagc
     1441 acgagctcgt gttgaatgtt tcgacaagtt taaggtcaac agcacactag aacaatacgt
     1501 cttctgcact gttaatgctt tgccagaaac gactgctgat atcgtcgtct ttgatgaaat
     1561 ctcgatggcc actaactatg acttatccgt agttaatgct agacttcgtg caaagcacta
     1621 cgtctatatt ggtgatccag ctcagctgcc tgcaccaaga actttgttaa ccaagggtac
     1681 cttggaacca gaatacttca actctgtgtg tcgcctgatg aagacaatcg gccctgatat
     1741 gttcctaggt acatgtagac gttgtccagc tgaaattgtg gacacggtca gtgccttggt
     1801 ttacgataac aaattaaagg cgcataagga caaaagcgct caatgtttca agatgtttta
     1861 caagggtgtc attactcacg atgttagttc ggccatcaat agaccccaaa taggtgttgt
     1921 tagagaattt ttgacaagga acccagcatg gagaaaggct gttttcatat ccccatacaa
     1981 ttctcaaaac gcggtagcct caaaaatctt gggtcttccc actcagaccg ttgattcttc
     2041 ccaaggaagt gaatatgact acgtgatttt cactcaaacc accgagactg cccactcttg
     2101 caacgtcaat cgtttcaacg ttgctattac gagagccaag gttggcatct tgtgtatcat
     2161 gtctgataga gacttgtacg ataaattaca gtttacatcc ttagaaatac caagacgaaa
     2221 cgtcgctact ttgcaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat
     2281 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
     2341 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
     2401 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg
     2461 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     2521 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     2581 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
     2641 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     2701 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     2761 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     2821 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     2881 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt
     2941 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     3001 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     3061 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     3121 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     3181 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     3241 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     3301 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     3361 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     3421 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     3481 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga
     3541 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     3601 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     3661 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     3721 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     3781 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     3841 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     3901 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     3961 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     4021 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     4081 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     4141 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     4201 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     4261 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     4321 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     4381 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     4441 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     4501 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct
     4561 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     4621 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4741 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5101 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc
     5161 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     5221 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     5281 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     5341 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     5401 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     5461 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct
     5521 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5581 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5641 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg
     5701 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5761 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5821 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5881 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca
     5941 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6001 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6061 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6121 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac
     6181 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6241 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6301 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct
     6361 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     6421 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     6481 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga
//
