LOCUS       sequence-295631-        8165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian 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 8165)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8165)
  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-m4133997
FEATURES             Location/Qualifiers
     source          1..8165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1589..1609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1610..1636
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1705..1826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1716..1735)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1770..1789
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1941..1987
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2017..2153
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2198..2269
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2495..3083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2575..2594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3583..4792
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4629..4648
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4796..5824
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5851..5899
                     /note="synthetic polyadenylation signal"
     CDS             join(6165..6233,6234..7025)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6383..6402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7038..7174
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7319..7390
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7395..7441)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7459..7507
                     /note="synthetic polyadenylation signal"
     enhancer        7588..7891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7893..8095
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8046..8066
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8164..8165,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcc gaaaatgtca ctggcttatt taaggattgc agtaaagtga tcacagggtt
       61 gcacccgacc caagcgccta cgcatctttc agtagacacc aagttcaaaa ctgagggact
      121 ctgtgtggac ataccaggta ttcccaagga tatgacatac agacgcctga tctcgatgat
      181 gggcttcaag atgaactatc aggttaacgg ttaccccaat atgtttatta cccgagagga
      241 agctatccgg cacgtgcgtg cctggatagg gtttgacgtc gagggctgtc atgcaactag
      301 ggaagctgta ggaacaaatc tgccactgca actcggcttc tctaccggag tcaacctagt
      361 tgccgtgcct accgggtatg tggatacacc aaacaatact gattttagca gagtctccgc
      421 aaaaccccct cccggggacc agttcaaaca cttgatcccc ctgatgtaca agggtttacc
      481 atggaacgtg gtgcggatta aaatagttca gatgctcagc gacacactga agaatttaag
      541 tgatagagtt gtgtttgtac tttgggccca tggcttcgag ctgacctcta tgaagtattt
      601 cgttaaaatt ggacccgaac gaacgtgctg cctgtgtgac cggagggcga cgtgtttctc
      661 aactgcttcc gatacctatg cctgctggca ccatagcatc ggctttgatt acgtgtacaa
      721 cccttttatg attgacgtgc agcagtgggg attcactggg aacctacagt cgaatcacga
      781 tctttactgt caggtccatg ggaacgcaca cgttgctagc tgcgacgcaa tcatgacacg
      841 ttgccttgct gtccacgagt gttttgtgaa acgagtggac tggactattg agtatcctat
      901 cattggcgac gaactgaaga taaatgcggc ttgtaggaag gtgcagcata tggtcgtcaa
      961 agccgccctc ttagctgata agttcccggt cctgcacgat attggcaatc caaaggccat
     1021 caaatgcgta ccccaggcag atgttgagtg gaagttttac gacgcgcaac catgctccga
     1081 taaggcctat aaaatcgaag agctcttcta cagttatgcc acccattctg acaagtttac
     1141 cgacggtgtg tgtttgtttt ggaattgcaa cgtggatcgc taccctgcaa actctatcgt
     1201 ttgtcgcttc gacacacgcg tattgtccaa tctgaatcta cccggatgcg acggaggttc
     1261 actgtatgtg aacaagcacg ccttccacac ccccgcattc gataaatccg cctttgtcaa
     1321 ccttaaacaa ttacctttct tttactacag cgactctcct tgcgaatccc atggaaagca
     1381 agtagtgagt gatatcgatt acgtcccact caaaagtgca acatgtatta cgagatgcaa
     1441 tctcggtggt gcagtctgta gacaccatgc caatgaatat cggctgtatt tggacgccta
     1501 taacatgatg atttcagctg gcttctctct ttgggtttac aagcagtttg acacgtataa
     1561 tctgtggaac acctttacaa ggctgcaaga aaacctttac tttcagtcgt acccctatga
     1621 cgtaccagat tatgcgtaag gatccgcagg cctctgctag cttgactgac tgagatacag
     1681 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1741 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1801 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1861 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     1921 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1981 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     2041 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     2101 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     2161 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     2221 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     2281 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     2341 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     2401 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     2461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3001 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3061 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3121 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3181 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3241 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3301 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     3361 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     3421 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     3481 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     3541 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     3601 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     3661 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     3721 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     3781 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     3841 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     3901 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3961 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     4021 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     4081 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     4141 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     4201 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     4261 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     4321 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     4381 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     4441 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     4501 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     4561 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     4621 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     4681 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     4741 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     4801 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4861 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4921 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4981 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     5041 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     5101 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     5161 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     5221 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     5281 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5341 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5401 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5461 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5521 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     5581 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5641 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     5701 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     5761 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     5821 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     5881 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     5941 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     6001 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     6061 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     6121 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     6181 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6241 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     6301 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     6361 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6421 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     6481 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6541 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     6601 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6661 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6721 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     6781 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6841 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6901 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6961 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7021 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     7081 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     7141 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     7201 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     7261 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     7321 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     7381 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     7441 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     7501 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     7561 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     7621 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     7681 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7741 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     7801 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     7861 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     7921 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     7981 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     8041 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     8101 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     8161 gccgc
//
