LOCUS       sequence-300421-        7414 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 7414)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7414)
  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-m4046289
FEATURES             Location/Qualifiers
     source          1..7414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(331..1134)
                     /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"
     primer_bind     1485..1503
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2646..2663)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(4255..4668)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4714..4745
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        4746..4848
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4849..4917,4918..5709)
                     /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     complement(5067..5086)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             5710..5901
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      5925..5952
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5964..6551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6573..6602)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(6680..7414,1..248)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt
       61 ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt
      121 cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc
      181 tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat
      241 gcttcgttac tcgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
      301 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
      361 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
      421 aaatagtcct cttccaacaa taataatgtc agatcctgta gaaaccacat catccacggt
      481 tctatactgt tgacccaatg catctccctt gtcatctaaa cccacaccgg gtgtcataat
      541 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
      601 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
      661 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
      721 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
      781 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
      841 accaatgtca gcaaattttc tgtcctcgaa gagtaaaaaa ttgtacttgg cggataatgc
      901 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
      961 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     1021 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     1081 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     1141 tcttcgtttc ctgcacgttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     1201 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     1261 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gtgtactgca
     1321 gtgcactagt ggttcggcgg ccgctttgta cagaaaaaaa agaaaaattt gaaatataaa
     1381 taacgttctt aatactaaca taactattaa aaaaaataaa tagggaccta gacttcaggt
     1441 tgtctaactc cttccttttc ggttagagcg gatgtgggag gagggcgtga atgtaagcgt
     1501 gacataacta attacatgat atcgacaaag gaaaaggggc ctgtttacgt ttgagcatct
     1561 tttaatgctt cgtcgacggt ttgtctaccg gaagcagcat taatcacggc agtcctaact
     1621 gcgtaccaga aggctgacat ttgtgggatg tttggcataa tttcaccctt ttgggcgttc
     1681 tccatggtag cggcaattct gggatcctta gccaattctt cttcgtaaga tttcaaagcg
     1741 acagcaccca atggcttgtc cttgttaacg gcctccaaac cttcgtcagt taacaagtaa
     1801 ttttccaaaa attctttagc tagctccttg tttggagaag cagcattgat gccagcggat
     1861 agaacaccga caaagggctt ggatggttgg cctttgaatg taggcaaaac agtgacacca
     1921 tagtttactt tagaagtatc aatgttactc caggcccatg gaccatttat ggtcatagca
     1981 gtttcaccct tgttgaaagc ggcttcggcg atagagtagt cagtgtcagc gttcatgtgt
     2041 ttattcttta ttaaatcaac caagaaggtc aaaccagcct ttgcaccagc attatcaacg
     2101 ccgacatcct taatgtcata cttaccgttt tcgtatttaa atgcgtaacc accgtcagca
     2161 gcgattaatg gccaggtgaa ataaggttct tgtaagttaa acatcaaagc gctttttccc
     2221 ttggccttta gctctttatc caaagctgga atttcttccc atgtctttgg tgggttaggt
     2281 aacaagtcct tattgtagat caacgataag gcttcaacag caattggata tgctataagc
     2341 ttaccgttgt atctgacagc atcccaagtg aatgggtaca atttatcttg gaaggccttg
     2401 tctggggtga tctcagccag caaacctgac tgagcatatc caccaaaacg atcgtgggcc
     2461 cagaaaataa tgtcgggacc atcaccagta gcagcaactt gagggaattt ttcttctaac
     2521 ttgtctggat gttcaaccgt caccttgata cctgtgtcct tttcaaattt cttgcctact
     2581 tctgccaaac cgttgtaacc cttatctcca tttatccaaa tgactagctt accttcctcg
     2641 attttagaac ctctcggcac gagaagtctt gtaaaagtgt tccagaggtt ataagtatca
     2701 aattgtttgt aaacccacaa gctaaagcca gctgagatca tcatgttata agcatcgaga
     2761 tacaatctgt actcattagc atgatgtcta cagacagcac cacctaaatt gcaacgtgtt
     2821 atacacgtag cagactttag tggtacataa tctatatctg acactacttg ttttccatga
     2881 gactcacatg gactgtcaga gtaatagaaa aatggtaatt gttttaaatt aacaaaagca
     2941 cttttatcaa aagctggtgt gtggaatgca tgtttattta catacaaact gccaccatca
     3001 caaccaggca agttaaggtt agatagcact ctagtgtcaa atctacaaac aatggaatta
     3061 gcaggatatc tatcgacatt gcaattccaa aataggcata caccatctgt gaatttgtca
     3121 gaatgtgtgg cataagaata gaataattct tctattttat aagctttgtc actacaaggc
     3181 tgtgcatcat agaacttcca ttctacatca gcttgaggta cacacttaat agctttaggg
     3241 ttaccaatgt cgtgaagaac tgggaatttg tctgctaata atgcagcttt aacaaccatg
     3301 tgttgaacct ttctacaagc cgcattaatc ttcagttcat caccaattat aggatattca
     3361 atagtccagt caacacgctt aacaaagcac tcgtggacag ctagacacct agtcatgatt
     3421 gcatcacaac tagctacatg tgcattacca tggacttgac aatacagatc atggttgctt
     3481 tgtaggttac ctgtaaaacc ccattgttga acatcaatca taaacggatt atagacgtaa
     3541 tcaaatccaa tagaatgatg ccaacaggca taagtgtctg aagcagtgga aaagcatgtg
     3601 gcacgtctat cacatagaca acaggtgcgc tcaggtccta ttttcacaaa atacttcata
     3661 gatgtcaact caaagccatg tgcccataag acaaatacga ctctgtcaga gagattttta
     3721 agtgtgtcac ttaacatttg tacaatcttt atacgcacta cattccaagg aagtcctttg
     3781 tacataagtg gtatgaggtg tttaaattga tctccaggcg gtggtttagc actaactctg
     3841 gaaaaatctg tattattagg tgtatcaaca taacctgtag gtacagcaac taggttaaca
     3901 cctgtagaaa aacctagctg taaaggtaaa ttggtaccaa cagcttctct agtagcatga
     3961 cacccctcga catcgaagcc aatccatgca cgtacatgtc ttatagcttc ttcgcgggtg
     4021 ataaacatgt tagggtaacc attaacttga taattcattt taaaacccat catagagatg
     4081 agtcttctat aggtcatgtc cttaggtatg ccaggtatgt caacacataa accttcagtt
     4141 ttgaatttag tgtcaacact gaggtgtgta ggtgcctgtg taggatgtaa cccagtgatt
     4201 accttactac aatctttaaa gagtcctgtt acattttcca ttttgtaatt aaaacttaga
     4261 ttagattgct atgctttctt tctaatgagc aagaagtaaa aaaagttgta atagaacaag
     4321 aaaaatgaaa ctgaaacttg agaaattgaa gaccgtttat taacttaaat atcaatggga
     4381 ggtcatcgaa agagaaaaaa atcaaaaaaa aaaattttca agaaaaagaa acgtgataaa
     4441 aatttttatt gcctttttcg acgaagaaaa agaaacgagg cggtctcttt tttcttttcc
     4501 aaacctttag tacgggtaat taacgacacc ctagaggaag aaagagggga aatttagtat
     4561 gctgtgcttg ggtgttttga agtggtacgg cgatgcgcgg agtccgagaa aatctggaag
     4621 agtaaaaaag gagtagaaac attttgaagc tatggtgtgt ggggcggccg ctctaactga
     4681 ccatttaaat catacctgac ctccatagca gaaagtcaaa agcctccgac cggaggcttt
     4741 tgacttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     4801 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat gagtattcaa
     4861 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4981 atcgaactgg atctcaacag cggtaagatc cttgagagtt tacgccccga agaacgtttt
     5041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5161 ccagtcacag aaaagcatct cacggatggc atgacagtaa gagaattatg cagtgctgcc
     5221 ataaccatga gtgataacac tgcggccaac ttacttctgg caacgatcgg aggaccgaag
     5281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5461 ttaatagact ggatggaggc ggataaagtt gcaggatcac ttctgcgctc ggccctcccg
     5521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5581 gcagcactgg ggccagatgg taagccctcc cgcatcgtag ttatctacac gacggggagt
     5641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5701 cattggtaag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     5761 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     5821 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
     5881 gatgacggtg aaaacctctg atgagggccc aaatgtaatc acctggctca ccttcgggtg
     5941 ggcctttctg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6001 aatcgatgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6061 acccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6121 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6181 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6301 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6361 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6421 tgcgctctgc tgaagccagt tacctcggaa aaagagttgg tagctcttga tccggcaaac
     6481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6541 aaggatctca agaagatcct ttgattttct accgaagaaa ggcccacccg tgaaggtgag
     6601 ccagtgagtt gattgcagtc cagttacgct ggagtcaagc ttgtaacacg cggccgctcg
     6661 ggaagttcct attctctagc tagaaagtat aggaacttca gagcgctttt gaaaaccaaa
     6721 agcgctctga tgacgcactt tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta
     6781 aaatattgcg aataccgctt ccacaaacat tgctcaaaag tatctctttg ctatatatct
     6841 ctgtgctata tccctatata acctacccat ccacctttcg ctccttgaac ttgcatctaa
     6901 actcgacctc tacatttttt atgtttatct ctagtattac tctttagaca aaaaaattgt
     6961 agtaagaact attcatagag tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac
     7021 gtagtatata gagacaaaat agaagaaacc gttcataatt ttctgaccaa tgaagaatca
     7081 tcaacgctat cactttctgt tcacaaagta tgcgcaatcc acatcggtat agaatataat
     7141 cggggatgcc tttatcttga aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg
     7201 ggaagtggag tcaggctttt tttatggaag agaaaataga caccaaagta gccttcttct
     7261 aaccttaacg gacctacagt gcaaaaagtt atcaagagac tgcattatag agcgcacaaa
     7321 ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata gcgctctcgg gatgcatttt
     7381 tgtagaacaa aaaagaagta tagattcttt gttg
//
