LOCUS       sequence-300444-        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"
     terminator      1020..1049
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1071..1658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1670..1697)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1721..1912)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1913..2704,2705..2773))
                     /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     2536..2555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2774..2876)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2877..2908)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2975..3416
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2975..3092
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3387..3410
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(5272..5290)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             5641..6444
                     /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      join(6527..6567,1..942)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
       61 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
      121 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
      181 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
      241 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
      301 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
      361 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
      421 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
      481 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
      541 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
      601 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
      661 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
      721 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
      781 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
      841 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
      901 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
      961 ccgagcggcc gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg
     1021 gctcaccttc acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct
     1081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1141 tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc
     1201 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1561 taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat
     1621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc
     1681 ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat
     1741 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     1801 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     1861 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat
     1921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1981 gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg
     2041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2101 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
     2161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2341 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
     2401 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
     2461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2581 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2761 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     2821 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
     2881 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
     2941 gtcagttaga gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga
     3001 cagccctccg acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga
     3061 aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt
     3121 ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg
     3181 aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt
     3241 aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc
     3301 ataaccactt taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt
     3361 cataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa
     3421 aactataatg gttggggctt gtgttctttg caattcacag acttcattaa gatgtggtgc
     3481 ttgcatacgt agaccattct tatgttgtaa atgctgttac gaccatgtca tatcaacatc
     3541 acataaatta gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac
     3601 agatgtgact caactttact taggaggtat gagctattat tgtaaatcac ataaaccacc
     3661 cattagtttt ccattgtgtg ctaatggaca agtttttggt ttatataaaa atacatgtgt
     3721 tggtagcgat aatgttactg actttaatgc aattgcaaca tgtgactgga caaatgctgg
     3781 tgattacatt ttagctaaca cctgtactga aagactcaag ctttttgcag cagaaacgct
     3841 caaagctact gaggagacat ttaaactgtc ttatggtatt gctactgtac gtgaagtgct
     3901 gtctgacaga gaattacatc tttcatggga agttggtaaa cctagaccac cacttaaccg
     3961 aaattatgtc tttactggtt atcgtgtaac taaaaacagt aaagtacaaa taggagagta
     4021 cacctttgaa aaaggtgact atggtgatgc tgttgtttac cgaggtacaa caacttacaa
     4081 attaaatgtt ggtgattatt ttgtgctgac atcacataca gtaatgccat taagtgcacc
     4141 tacactagtg ccacaagagc actatgttag aattactggc ttatacccaa cactcaatat
     4201 ctcagatgag ttttctagca atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc
     4261 tacactccag ggaccacctg gtactggtaa gagtcatttt gctattggcc tagctctcta
     4321 ctacccttct gctcgcatag tgtatacagc ttgctctcat gccgctgttg atgcactatg
     4381 tgagaaggca ttaaaatatt tgcctataga taaatgtagt agaattatac ctgcacgtgc
     4441 tcgtgtagag tgttttgata aattcaaagt gaattcaaca ttagaacagt atgtcttttg
     4501 tactgtaaat gcattgcctg agacgacagc agatatagtt gtctttgatg aaatttcaat
     4561 ggccacaaat tatgatttga gtgttgtcaa tgccagatta cgtgctaagc actatgtgta
     4621 cattggcgac cctgctcaat tacctgcacc acgcacattg ctaactaagg gcacactaga
     4681 accagaatat ttcaattcag tgtgtagact tatgaaaact ataggtccag acatgttcct
     4741 cggaacttgt cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt tggtttatga
     4801 taataagctt aaagcacata aagacaaatc agctcaatgc tttaaaatgt tttataaggg
     4861 tgttatcacg catgatgttt catctgcaat taacaggcca caaataggcg tggtaagaga
     4921 attccttaca cgtaaccctg cttggagaaa agctgtcttt atttcacctt ataattcaca
     4981 gaatgctgta gcctcaaaga ttttgggact accaactcaa actgttgatt catcacaggg
     5041 ctcagaatat gactatgtca tattcactca aaccactgaa acagctcact cttgtaatgt
     5101 aaacagattt aatgttgcta ttaccagagc aaaagtaggc atactttgca taatgtctga
     5161 tagagacctt tatgacaagt tgcaatttac aagtcttgaa attccacgta ggaatgtggc
     5221 aactttataa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     5281 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     5341 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     5401 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     5461 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     5521 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     5581 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     5641 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     5701 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     5761 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     5821 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     5881 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     5941 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     6001 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     6061 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     6121 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     6181 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     6241 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     6301 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     6361 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     6421 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     6481 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     6541 gcttcatttt gtagaacaaa aatgcaa
//
