LOCUS       sequence-295621-        8528 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 8528)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8528)
  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-m4134105
FEATURES             Location/Qualifiers
     source          1..8528
                     /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             1958..1975
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2068..2189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2079..2098)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2133..2152
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2304..2350
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2380..2516
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2561..2632
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2858..3446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2938..2957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3946..5155
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4992..5011
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5159..6187
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6214..6262
                     /note="synthetic polyadenylation signal"
     CDS             join(6528..6596,6597..7388)
                     /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(6746..6765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7401..7537
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7682..7753
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7758..7804)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7822..7870
                     /note="synthetic polyadenylation signal"
     enhancer        7951..8254
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8256..8458
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8409..8429
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8527..8528,1..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 tttcagtcga tgagtgacaa
     1621 aatcattcat ctaaccgatg actcctttga tacagacgtg ttgaaggccg atggtgcgat
     1681 actggtagat ttctgggctg aatggtgtgg cccgtgcaag atgatcgcac ccattctgga
     1741 cgagatcgcc gacgagtatc aaggaaaatt aactgtcgct aagcttaata tagatcagaa
     1801 cccagggacg gcacctaaat acggcatcag gggaatacct accctcctgc tctttaagaa
     1861 tggggaagtt gccgcgacaa aagtgggagc cctgtctaag ggccagctaa aagagttctt
     1921 ggacgctaac ctggcaggtt caggctcggg gcacatgcac catcaccatc accatagcag
     1981 cgggctcgtg ccacgcggct aaggatccgc aggcctctgc tagcttgact gactgagata
     2041 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
     2101 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2161 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2221 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2281 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
     2341 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
     2401 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     2461 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
     2521 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
     2581 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
     2641 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
     2701 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     2761 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     2821 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2881 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3061 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3361 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3661 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     3721 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     3781 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     3841 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     3901 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     3961 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     4021 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     4081 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     4141 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     4201 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     4261 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     4321 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     4381 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     4441 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     4501 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     4561 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     4621 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     4681 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     4741 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     4801 gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg
     4861 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     4921 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     4981 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     5041 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     5101 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     5161 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     5221 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     5281 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     5341 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     5401 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     5461 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     5521 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     5581 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     5641 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     5701 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     5761 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     5821 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     5881 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     5941 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     6001 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     6061 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     6121 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     6181 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     6241 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     6301 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     6361 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     6421 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     6481 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     6541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     6661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     6721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     6841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6901 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     6961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7081 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     7141 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7381 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     7441 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     7501 tccatacgtg tcctccttac cagaaattta tccttaagat cgtttaaact cgactctggc
     7561 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     7621 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     7681 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     7741 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     7801 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     7861 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     7921 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     7981 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     8041 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     8101 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     8161 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     8221 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
     8281 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     8341 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     8401 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     8461 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
     8521 gcggccgc
//
