LOCUS       sequence-295767-        8174 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 8174)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8174)
  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-m4133788
FEATURES             Location/Qualifiers
     source          1..8174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1583..1603
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1604..1645
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1714..1835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1725..1744)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1779..1798
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1950..1996
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2026..2162
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2207..2278
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2504..3092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2584..2603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3592..4801
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4638..4657
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4805..5833
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5860..5908
                     /note="synthetic polyadenylation signal"
     CDS             join(6174..6242,6243..7034)
                     /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(6392..6411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7047..7183
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7328..7399
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7404..7450)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7468..7516
                     /note="synthetic polyadenylation signal"
     enhancer        7597..7900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7902..8104
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8055..8075
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8173..8174,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggaa aatgtaacag gactctttaa agattgtagt aaggtaatca ctgggttaca
       61 tcctacacag gcacctacac acctcagtgt tgacactaaa ttcaaaactg aaggtttatg
      121 tgttgacata cctggcatac ctaaggacat gacctataga agactcatct ctatgatggg
      181 ttttaaaatg aattatcaag ttaatggtta ccctaacatg tttatcaccc gcgaagaagc
      241 tataagacat gtacgtgcat ggattggctt cgatgtcgag gggtgtcatg ctactagaga
      301 agctgttggt accaatttac ctttacagct aggtttttct acaggtgtta acctagttgc
      361 tgtacctaca ggttatgttg atacacctaa taatacagat ttttccagag ttagtgctaa
      421 accaccgcct ggagatcaat ttaaacacct cataccactt atgtacaaag gacttccttg
      481 gaatgtagtg cgtataaaga ttgtacaaat gttaagtgac acacttaaaa atctctctga
      541 cagagtcgta tttgtcttat gggcacatgg ctttgagttg acatctatga agtattttgt
      601 gaaaatagga cctgagcgca cctgttgtct atgtgataga cgtgccacat gcttttccac
      661 tgcttcagac acttatgcct gttggcatca ttctattgga tttgattacg tctataatcc
      721 gtttatgatt gatgttcaac aatggggttt tacaggtaac ctacaaagca accatgatct
      781 gtattgtcaa gtccatggta atgcacatgt agctagttgt gatgcaatca tgactaggtg
      841 tctagctgtc cacgagtgct ttgttaagcg tgttgactgg actattgaat atcctataat
      901 tggtgatgaa ctgaagatta atgcggcttg tagaaaggtt caacacatgg ttgttaaagc
      961 tgcattatta gcagacaaat tcccagttct tcacgacatt ggtaacccta aagctattaa
     1021 gtgtgtacct caagctgatg tagaatggaa gttctatgat gcacagcctt gtagtgacaa
     1081 agcttataaa atagaagaat tattctattc ttatgccaca cattctgaca aattcacaga
     1141 tggtgtatgc ctattttgga attgcaatgt cgatagatat cctgctaatt ccattgtttg
     1201 tagatttgac actagagtgc tatctaacct taacttgcct ggttgtgatg gtggcagttt
     1261 gtatgtaaat aaacatgcat tccacacacc agcttttgat aaaagtgctt ttgttaattt
     1321 aaaacaatta ccatttttct attactctga cagtccatgt gagtctcatg gaaaacaagt
     1381 agtgtcagat atagattatg taccactaaa gtctgctacg tgtataacac gttgcaattt
     1441 aggtggtgct gtctgtagac atcatgctaa tgagtacaga ttgtatctcg atgcttataa
     1501 catgatgatc tcagctggct ttagcttgtg ggtttacaaa caatttgata cttataacct
     1561 ctggaacact tttacaagac ttgaaaacct ttactttcag tcggggaaac ctatacccaa
     1621 tccacttctc ggcttagatt ctacctaagg atccgcaggc ctctgctagc ttgactgact
     1681 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
     1741 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1801 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1861 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1921 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     1981 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     2041 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     2101 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     2161 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     2221 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     2281 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     2341 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     2401 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     2461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2761 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2821 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2881 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2941 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3001 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3061 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3121 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3181 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3241 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3301 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     3361 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     3421 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     3481 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     3541 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     3601 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     3661 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     3721 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     3781 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     3841 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     3901 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3961 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     4021 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     4081 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     4141 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     4201 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     4261 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     4321 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     4381 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     4441 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     4501 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     4561 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     4621 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     4681 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     4741 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     4801 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4861 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4921 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4981 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     5041 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     5101 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     5161 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     5221 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     5281 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     5341 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     5401 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     5461 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     5521 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     5581 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     5641 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5701 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     5761 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5821 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     5881 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     5941 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     6001 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     6061 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     6121 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     6181 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6241 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     6301 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6361 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6421 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6481 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6541 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     6601 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6661 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6721 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     6781 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6841 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6901 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6961 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7021 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     7081 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     7141 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac
     7201 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     7261 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     7321 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     7381 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7441 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     7501 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7561 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     7621 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     7681 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     7741 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     7801 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     7861 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     7921 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     7981 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     8041 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     8101 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     8161 ccgccagcgg ccgc
//
