LOCUS       sequence-298235-        6340 bp    DNA     circular SYN 15-APR-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 6340)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6340)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046198
FEATURES             Location/Qualifiers
     source          1..6340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2015..2032
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2033..2050
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2095..2113)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2464..3267
                     /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      3350..4332
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4410..4439
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4461..5048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5060..5087)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5111..5302)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5303..6094,6095..6163))
                     /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     5926..5945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6164..6266)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6267..6298)
                     /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 atggccgaga atgtcacagg cctattcaag gattgctcga aagtgatcac
      481 cggtttacac cctacgcagg ctccaactca tttgtccgtt gacaccaagt ttaagactga
      541 aggattgtgt gttgacatac caggtattcc caaagatatg acttaccgta ggctgatttc
      601 tatgatgggt ttcaagatga actatcaagt taacggttac ccaaatatgt ttatcaccag
      661 agaagaagct attagacacg tacgtgcatg gatcggtttc gatgtcgaag gttgtcatgc
      721 tactagagaa gcggtcggta caaacttgcc attacaactt ggcttttcaa ccggtgttaa
      781 tttggttgct gttcctacag ggtatgtcga cactccaaac aacacggatt tctctcgagt
      841 gtccgccaag ccaccacctg gtgatcaatt caaacacttg atcccgctca tgtacaaggg
      901 tttgccttgg aacgttgtca gaattaaaat agtacaaatg ttatctgata ctttgaagaa
      961 tctgagtgac agagttgtct tcgtcctttg ggctcacggt tttgagctaa ccagcatgaa
     1021 gtactttgtg aagattggac cagaacggac ttgttgcttg tgtgacagaa gagccacttg
     1081 tttctccacc gctagcgata catatgcatg ttggcatcat tctattggtt tcgactacgt
     1141 ttacaacccc tttatgatcg acgttcaaca gtggggtttc accggtaatt tgcaatcgaa
     1201 ccacgacttg tattgtcaag ttcacggcaa tgctcacgtg gcttcatgcg atgctattat
     1261 gacaaggtgt ttagccgtcc atgaatgttt cgttaaaaga gtagactgga ctatcgaata
     1321 cccaattatc ggtgatgaac tcaagattaa cgctgcctgt agaaaggtcc aacatatggt
     1381 tgtaaaggca gctctattag ctgacaaatt tccagttttg cacgatatag ggaatccaaa
     1441 ggccataaag tgtgttccac aagctgacgt cgaatggaaa ttctacgatg cgcaaccctg
     1501 ctctgataaa gcttataaga tcgaggaatt gttttactct tacgctaccc actccgataa
     1561 gttcactgac ggtgtgtgtc ttttctggaa ctgcaatgtt gatagatatc cagcaaactc
     1621 catcgtttgt agattcgaca cgcgagtctt gagcaacttg aacttaccag gttgtgatgg
     1681 tggttccttg tacgtcaata aacatgcatt tcacactccc gctttcgaca agtctgcttt
     1741 tgttaattta aagcaattgc cttttttcta ttactctgat agtccatgcg aatctcatgg
     1801 taagcaagtc gtttccgaca ttgattacgt accactgaag tcagccacct gtataacaag
     1861 atgtaacctt ggaggtgctg tctgccgcca ccatgctaac gagtacagac tatacttgga
     1921 tgcctataac atgatgattt cagccggttt ctctttatgg gtttacaagc agtttgacac
     1981 ttacaacttg tggaacacct tcacgcgttt gcaactcgtg ccgagaggtt ctcaccatca
     2041 tcaccatcat taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     2101 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     2161 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     2221 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     2281 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     2341 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     2401 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     2461 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     2521 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     2581 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     2641 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     2701 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     2761 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     2821 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     2881 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     2941 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     3001 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     3061 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     3121 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     3181 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     3241 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     3301 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     3361 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     3421 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     3481 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     3541 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     3601 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     3661 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     3721 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     3781 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     3841 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     3901 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     3961 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     4021 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     4081 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     4141 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     4201 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     4261 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     4321 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     4381 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     4441 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4501 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4561 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4621 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4681 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4741 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4801 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4861 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4921 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     4981 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5041 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     5101 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     5161 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     5221 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     5281 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     5341 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     5401 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5461 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     5521 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5581 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5641 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5701 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     5761 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5821 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5881 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5941 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     6001 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6061 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6121 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     6181 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     6241 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     6301 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
