LOCUS       sequence-298126-        6347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6347)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6347)
  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-m4046224
FEATURES             Location/Qualifiers
     source          1..6347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..418
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2016..2033
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2034..2057
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2102..2120)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2471..3274
                     /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      3357..4339
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4417..4446
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4468..5055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5067..5094)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5118..5309)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5310..6101,6102..6170))
                     /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     5933..5952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6171..6273)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6274..6305)
                     /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 ttttttttng atttttttct ctttcgatga cctcccattg
      301 atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct
      361 attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt
      421 ttaattacaa aatggccgag aatgtcacag gcctattcaa ggattgctcg aaagtgatca
      481 ccggtttaca ccctacgcag gctccaactc atttgtccgt tgacaccaag tttaagactg
      541 aaggattgtg tgttgacata ccaggtattc ccaaagatat gacttaccgt aggctgattt
      601 ctatgatggg tttcaagatg aactatcaag ttaacggtta cccaaatatg tttatcacca
      661 gagaagaagc tattagacac gtacgtgcat ggatcggttt cgatgtcgaa ggttgtcatg
      721 ctactagaga agcggtcggt acaaacttgc cattacaact tggcttttca accggtgtta
      781 atttggttgc tgttcctaca gggtatgtcg acactccaaa caacacggat ttctctcgag
      841 tgtccgccaa gccaccacct ggtgatcaat tcaaacactt gatcccgctc atgtacaagg
      901 gtttgccttg gaacgttgtc agaattaaaa tagtacaaat gttatctgat actttgaaga
      961 atctgagtga cagagttgtc ttcgtccttt gggctcacgg ttttgagcta accagcatga
     1021 agtactttgt gaagattgga ccagaacgga cttgttgctt gtgtgacaga agagccactt
     1081 gtttctccac cgctagcgat acatatgcat gttggcatca ttctattggt ttcgactacg
     1141 tttacaaccc ctttatgatc gacgttcaac agtggggttt caccggtaat ttgcaatcga
     1201 accacgactt gtattgtcaa gttcacggca atgctcacgt ggcttcatgc gatgctatta
     1261 tgacaaggtg tttagccgtc catgaatgtt tcgttaaaag agtagactgg actatcgaat
     1321 acccaattat cggtgatgaa ctcaagatta acgctgcctg tagaaaggtc caacatatgg
     1381 ttgtaaaggc agctctatta gctgacaaat ttccagtttt gcacgatata gggaatccaa
     1441 aggccataaa gtgtgttcca caagctgacg tcgaatggaa attctacgat gcgcaaccct
     1501 gctctgataa agcttataag atcgaggaat tgttttactc ttacgctacc cactccgata
     1561 agttcactga cggtgtgtgt cttttctgga actgcaatgt tgatagatat ccagcaaact
     1621 ccatcgtttg tagattcgac acgcgagtct tgagcaactt gaacttacca ggttgtgatg
     1681 gtggttcctt gtacgtcaat aaacatgcat ttcacactcc cgctttcgac aagtctgctt
     1741 ttgttaattt aaagcaattg ccttttttct attactctga tagtccatgc gaatctcatg
     1801 gtaagcaagt cgtttccgac attgattacg taccactgaa gtcagccacc tgtataacaa
     1861 gatgtaacct tggaggtgct gtctgccgcc accatgctaa cgagtacaga ctatacttgg
     1921 atgcctataa catgatgatt tcagccggtt tctctttatg ggtttacaag cagtttgaca
     1981 cttacaactt gtggaacacc ttcacgcgtt tgcaactcgt gccgagaggt tcttggagcc
     2041 atccgcagtt tgaaaaataa acaggcccct tttcctttgt cgatatcatg taattagtta
     2101 tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta
     2161 gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac
     2221 gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt
     2281 gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga
     2341 agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga
     2401 aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg
     2461 aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt
     2521 tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga
     2581 tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt
     2641 actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct
     2701 aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat
     2761 tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga
     2821 cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc
     2881 agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg
     2941 ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga
     3001 ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg
     3061 gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta
     3121 tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact
     3181 atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga
     3241 agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg
     3301 tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg
     3361 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     3421 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     3481 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     3541 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     3601 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     3661 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     3721 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     3781 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     3841 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     3901 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     3961 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     4021 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     4081 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     4141 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     4201 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     4261 gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg
     4321 aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc
     4381 ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt
     4441 tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4561 aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4621 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4681 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4741 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4801 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4861 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4921 gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt
     4981 cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg
     5041 cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga
     5101 ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     5161 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     5221 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     5281 ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc
     5341 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5401 acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5461 tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt
     5521 gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5581 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5641 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5701 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc
     5761 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5821 actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc
     5881 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5941 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa
     6001 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6061 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6121 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc
     6181 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     6241 tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt
     6301 tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
