LOCUS       sequence-295639-        6681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6681)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6681)
  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-m4134539
FEATURES             Location/Qualifiers
     source          1..6681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1922..1942
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1976..2053
                     /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             2081..2101
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2550..2671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2561..2580)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2615..2634
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2786..2832
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2862..2998
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3043..3114
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3340..3928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3420..3439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4674..4742,4743..5534)
                     /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(4892..4911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5547..5683
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5835..5906
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5911..5957)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5975..6023
                     /note="synthetic polyadenylation signal"
     enhancer        6104..6407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6409..6611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6562..6582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6680..6681,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcc tacacaagat atgtggataa caatttttgt gggccggacg gatacccttt
       61 agaatgcatc aaagatttgc ttgcaagggc gggcaaggct tcgtgcactc tgtccgagca
      121 gctggacttc attgatacca agcgcggtgt ttattgttgc cgagagcatg aacacgagat
      181 agcctggtat acggaacggt cagagaaaag ttacgaactc caaaccccat tcgagatcaa
      241 gctagctaaa aagtttgaca ctttcaacgg cgaatgtccc aattttgtct ttcccctgaa
      301 ctctattatc aaaacaatac agcctcgggt ggagaagaag aaattagacg gattcatggg
      361 gcgtattagg agcgtatacc cagtggccag cccgaatgaa tgcaaccaga tgtgtctttc
      421 cacactgatg aagtgcgatc actgtggcga gacttcttgg caaaccggtg acttcgtaaa
      481 agcgacatgc gagttttgtg gtaccgagaa tctgactaag gagggagcta ccacatgcgg
      541 ctatttgcca cagaatgcag tggtcaagat ctactgtcct gcatgccata actctgaagt
      601 cggccccgag cactcgctcg ccgaatatca caatgagagc gggctaaaaa ctatactgag
      661 gaaaggaggt cgcaccatcg cctttggggg ctgcgttttc tcctacgtgg ggtgtcataa
      721 caaatgtgcc tattgggttc ccagagcctc cgctaacatt gggtgcaatc atacgggagt
      781 ggtcggggaa ggctcagaag gcttgaacga taacctcctt gaaattctgc aaaaggagaa
      841 ggtgaatatc aatatcgttg gcgacttcaa gttaaatgag gaaatcgcga ttattctggc
      901 aagtttttct gcctcaacta gcgcctttgt agagacagtg aaaggactcg attataaagc
      961 tttcaaacag atagtcgaga gttgtggtaa cttcaaggtt acaaagggca aggctaaaaa
     1021 gggtgcttgg aacattggag agcagaagtc aatcttgagt ccactgtacg cgtttgcatc
     1081 tgaggccgcc cgtgtggtca ggagcatatt cagtcgaacc ctagaaaccg cacagaatag
     1141 cgtgcgggta ttacagaagg cggctattac tattctcgac ggtatctcgc agtattctct
     1201 gagactgatc gacgccatga tgtttacaag cgatctggca acgaataact tagtggtcat
     1261 ggcgtacata accggagggg tcgtgcaact tacctcccag tggctaacga acatctttgg
     1321 gacggtttac gagaagctaa aacctgtgct ggactggctc gaagagaaat tcaaagaagg
     1381 cgtggaattt ttgcgtgacg ggtgggagat cgtaaagttc atatcaacat gtgcttgcga
     1441 aatcgttggc ggacaaatcg taacttgcgc aaaagagatt aaagaatcgg tgcagacctt
     1501 ctttaaactt gttaacaaat tcctggcttt gtgtgccgat tctattatca tcgggggagc
     1561 aaagcttaaa gccctaaatt taggcgagac atttgtgacg cacagtaagg gcctgtatag
     1621 aaagtgcgtg aagagtcgcg aagaaaccgg actgctcatg cccttaaaag cgccaaaaga
     1681 aataattttc ctggaagggg agacattgcc cactgaagtg ctcactgagg aagtagttct
     1741 gaaaactggg gatctgcaac ctttagagca gccgacctcc gaggccgtcg aggcacctct
     1801 cgtgggaacc ccggtctgta ttaatgggtt gatgctgctc gagattaagg ataccgaaaa
     1861 atactgcgca ctagccccca acatgatggt gactaacaac acatttaccc tcaagggagg
     1921 ggaaaacctt tactttcagt cgggacggcg aatcccgggg ctgataaatc cctggaaaag
     1981 gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc
     2041 aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct atttccaggg
     2101 tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata aagaacagca
     2161 gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac gcaacgcttt
     2221 cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg aagccaagaa
     2281 gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc agcagaatgc
     2341 tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg catttatcca
     2401 gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca aaaagctcaa
     2461 tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga ctgactgaga
     2521 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     2581 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2641 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2701 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2761 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     2821 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     2881 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     2941 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     3001 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     3061 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     3121 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     3181 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     3241 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     3301 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3361 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3421 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3481 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3541 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3601 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3661 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3721 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3781 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3841 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3901 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3961 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4021 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4081 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4141 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     4201 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     4261 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     4321 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     4381 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     4441 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     4501 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     4561 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     4621 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     4681 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4741 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     4801 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4861 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4921 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4981 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5041 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     5101 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5161 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5221 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     5281 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5341 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5401 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5461 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5521 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     5581 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     5641 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     5701 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     5761 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     5821 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     5881 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     5941 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     6001 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     6061 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     6121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     6181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     6241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     6301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     6361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     6421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     6481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     6541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     6601 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     6661 gacacacccg ccagcggccg c
//
