LOCUS       sequence-295267-        8183 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 8183)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8183)
  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-m4134103
FEATURES             Location/Qualifiers
     source          1..8183
                     /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..1654
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1723..1844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1734..1753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1788..1807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1959..2005
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2035..2171
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2216..2287
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2513..3101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2593..2612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3601..4810
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4647..4666
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4814..5842
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5869..5917
                     /note="synthetic polyadenylation signal"
     CDS             join(6183..6251,6252..7043)
                     /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(6401..6420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7056..7192
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7337..7408
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7413..7459)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7477..7525
                     /note="synthetic polyadenylation signal"
     enhancer        7606..7909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7911..8113
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8064..8084
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8182..8183,1..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 aagaaaccgc
     1621 ggcagctaag ttcgagaggc agcacatgga cagttaagga tccgcaggcc tctgctagct
     1681 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
     1741 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1801 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1861 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1921 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
     1981 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
     2041 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     2101 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     2161 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     2221 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     2281 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     2341 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     2401 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     2461 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2521 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2581 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2641 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2701 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2761 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2821 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2881 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2941 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3001 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3061 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3301 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     3361 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     3421 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     3481 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     3541 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     3601 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     3661 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     3721 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     3781 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     3841 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     3901 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     3961 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     4021 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     4081 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     4141 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     4201 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     4261 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     4321 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     4381 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     4441 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     4501 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     4561 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     4621 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     4681 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     4741 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     4801 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     4861 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     4921 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     4981 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     5041 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     5101 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     5161 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     5221 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     5281 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     5341 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     5401 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     5461 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     5521 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     5581 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     5641 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     5701 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     5761 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     5821 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     5881 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     5941 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     6001 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     6061 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     6121 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     6181 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6241 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6301 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6361 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6421 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6481 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6541 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6601 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6661 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6721 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     6781 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     6841 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6901 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6961 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7021 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     7081 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     7141 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt
     7201 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     7261 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     7321 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     7381 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     7441 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     7501 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     7561 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     7621 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7681 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7741 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     7801 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     7861 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     7921 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     7981 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     8041 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     8101 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     8161 tcgacacacc cgccagcggc cgc
//
