LOCUS       sequence-295846-        8693 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 8693)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8693)
  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-m4133523
FEATURES             Location/Qualifiers
     source          1..8693
                     /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             1643..1720
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1748..1768
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2233..2354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2244..2263)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2298..2317
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2469..2515
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2545..2681
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2726..2797
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3023..3611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3103..3122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4111..5320
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5157..5176
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5324..6352
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6379..6427
                     /note="synthetic polyadenylation signal"
     CDS             join(6693..6761,6762..7553)
                     /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(6911..6930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7566..7702
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7847..7918
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7923..7969)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7987..8035
                     /note="synthetic polyadenylation signal"
     enhancer        8116..8419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8421..8623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8574..8594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8692..8693,1..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 gacggcgaat
     1621 cccggggctg ataaatccct ggaaaaggag atggaagaag aactttattg cagtgtctgc
     1681 ggctaaccgg ttcaaaaaga tcagttcaag cggcgcccta gattacgaca ttcctacgac
     1741 cgcctccgag aatctctatt tccagggtga atttggcttg gcacaacatg acgaggccgt
     1801 cgataacaag tttaataaag aacagcagaa cgctttctat gagatccttc acttaccaaa
     1861 tctgaacgaa gagcaacgca acgctttcat tcagagcctg aaagacgatc cctcgcagtc
     1921 tgcaaatctc ctggccgaag ccaagaagct gaatgacgca caggctccaa aagttgataa
     1981 caagtttaac aaggagcagc agaatgcttt ctacgagata ttgcaccttc ctaatctgaa
     2041 tgaggaacaa agaaatgcat ttatccagtc cctaaaagac gatccatcac agagtgccaa
     2101 cctgcttgcg gaagccaaaa agctcaatga tgcccaggcc ccgaaagtag acgcgaacca
     2161 tcaataagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
     2221 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2281 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2341 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2401 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
     2461 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
     2521 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
     2581 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     2641 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
     2701 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
     2761 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
     2821 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     2881 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     2941 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3841 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     3901 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     3961 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     4021 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     4081 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     4141 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     4201 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     4261 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     4321 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     4381 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     4441 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     4501 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     4561 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     4621 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     4681 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     4741 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     4801 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     4861 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     4921 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     4981 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     5041 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     5101 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     5161 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     5221 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     5281 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     5341 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     5401 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     5461 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     5521 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     5581 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     5641 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     5701 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     5761 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     5821 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     5881 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     5941 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     6001 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     6061 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     6121 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     6181 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     6241 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     6301 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     6361 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     6421 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     6481 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     6541 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     6601 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     6661 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     6721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6781 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     6841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     6901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6961 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     7021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7081 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     7141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     7261 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     7321 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     7561 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     7621 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     7681 cttaccagaa atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg
     7741 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     7801 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     7861 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     7921 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     7981 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     8041 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     8101 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     8161 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     8221 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     8281 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     8341 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     8401 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     8461 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     8521 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     8581 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     8641 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
