LOCUS       sequence-295826-        8210 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 8210)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8210)
  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-m4133418
FEATURES             Location/Qualifiers
     source          1..8210
                     /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..1681
                     /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"
     polyA_signal    1750..1871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1761..1780)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1815..1834
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1986..2032
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2062..2198
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2243..2314
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2540..3128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2620..2639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3628..4837
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4674..4693
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4841..5869
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5896..5944
                     /note="synthetic polyadenylation signal"
     CDS             join(6210..6278,6279..7070)
                     /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(6428..6447)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7083..7219
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7364..7435
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7440..7486)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7504..7552
                     /note="synthetic polyadenylation signal"
     enhancer        7633..7936
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7938..8140
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8091..8111
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8209..8210,1..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 tcgaagaggc gatggaaaaa
     1621 gaattttatc gcggtgagtg cagccaacag attcaaaaag atatcctcgt ctggtgccct
     1681 ctaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     1741 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1801 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1861 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1921 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     1981 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     2041 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     2101 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     2161 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     2221 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     2281 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     2341 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     2401 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     2461 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2701 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2941 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3061 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3181 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3241 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3301 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3361 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     3421 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     3481 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     3541 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     3601 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     3661 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     3721 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     3781 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     3841 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     3901 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     3961 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     4021 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     4081 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     4141 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     4201 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     4261 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     4321 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     4381 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     4441 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     4501 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     4561 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     4621 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     4681 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     4741 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     4801 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     4861 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4921 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4981 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     5041 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     5101 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     5161 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     5221 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5281 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     5341 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     5401 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     5461 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     5521 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     5581 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     5641 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     5701 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5761 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     5821 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     5881 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     5941 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     6001 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     6061 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     6121 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6181 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6241 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6301 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6361 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6421 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6481 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     6541 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6601 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6661 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6721 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6781 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6841 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6901 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6961 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7021 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     7081 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     7141 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     7201 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     7261 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     7321 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     7381 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     7441 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     7501 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     7561 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     7621 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7681 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7741 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7801 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7861 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7921 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7981 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8041 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     8101 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     8161 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
