LOCUS       sequence-307280-        8187 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 8187)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8187)
  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-m4134022
FEATURES             Location/Qualifiers
     source          1..8187
                     /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..1651
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1720..1841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1731..1750)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1785..1804
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1956..2002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2032..2168
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2213..2284
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2510..3098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2590..2609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3598..4807
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4644..4663
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4811..5839
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5866..5914
                     /note="synthetic polyadenylation signal"
     CDS             join(6180..6248,6249..7040)
                     /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(6398..6417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7053..7189
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7341..7412
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7417..7463)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7481..7529
                     /note="synthetic polyadenylation signal"
     enhancer        7610..7913
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7915..8117
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8068..8088
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8186..8187,1..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 tttcagtcgg ggaaacctat
     1621 acccaatcca cttctcggct tagattctac ctaaggatcc gcaggcctct gctagcttga
     1681 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
     1741 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1801 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1861 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1921 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
     1981 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
     2041 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     2101 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     2161 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     2221 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     2281 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     2341 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     2401 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     2461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3301 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     3361 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     3421 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     3481 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     3541 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc
     3601 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca
     3661 cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     3721 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     3781 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     3841 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     3901 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     3961 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     4021 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     4081 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     4141 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     4201 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct
     4261 aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt
     4321 gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg
     4381 cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac
     4441 gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta
     4501 ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca
     4561 ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt
     4621 atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc
     4681 gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg
     4741 taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt
     4801 gttagacaac atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     4861 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4921 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4981 tttctacaaa gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga
     5041 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     5101 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     5161 ggaggcaatg gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt
     5221 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     5281 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     5341 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     5401 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     5461 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     5521 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     5581 tgcaggatcg ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     5641 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     5701 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt
     5761 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     5821 tccgagggca aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc
     5881 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     5941 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     6001 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     6061 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     6121 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     6181 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6241 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     6301 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6361 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6421 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6481 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6541 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     6601 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6661 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6721 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     6781 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6841 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6901 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6961 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7021 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     7081 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     7141 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     7201 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     7261 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     7321 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     7381 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     7441 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     7501 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     7561 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     7621 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     7681 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     7741 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     7801 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     7861 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     7921 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     7981 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     8041 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     8101 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     8161 ccactcgaca cacccgccag cggccgc
//
