LOCUS       sequence-295649-        8104 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 8104)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8104)
  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-m4134509
FEATURES             Location/Qualifiers
     source          1..8104
                     /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             1943..2020
                     /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    2073..2194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2084..2103)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2138..2157
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2309..2355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2385..2521
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2566..2637
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2863..3451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2943..2962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3951..5160
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4997..5016
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5164..5763
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5164..5183)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5660..5680
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5790..5838
                     /note="synthetic polyadenylation signal"
     CDS             join(6104..6172,6173..6964)
                     /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(6322..6341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6977..7113
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7258..7329
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7334..7380)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7398..7446
                     /note="synthetic polyadenylation signal"
     enhancer        7527..7830
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7832..8034
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7985..8005
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8103..8104,1..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 cgaagaggcg atggaaaaag aattttatcg cggtgagtgc
     1981 agccaacaga ttcaaaaaga tatcctcgtc tggtgccctc taagctagct tgactgactg
     2041 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     2101 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2161 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2221 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2281 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     2341 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     2401 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     2461 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     2521 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     2581 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     2641 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     2701 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     2761 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     2821 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2881 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2941 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3001 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3061 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3121 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3181 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3241 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3301 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3361 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3421 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3481 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3541 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3601 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3661 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     3721 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     3781 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     3841 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     3901 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg
     3961 ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga
     4021 cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc
     4081 tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg
     4141 ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc
     4201 cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag
     4261 gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg
     4321 tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt
     4381 tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta
     4441 gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg
     4501 cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa
     4561 caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg
     4621 gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt
     4681 gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc
     4741 gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc
     4801 gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt
     4861 ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg
     4921 gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc
     4981 tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg
     5041 ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt
     5101 cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac
     5161 aacatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     5221 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     5281 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5341 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5401 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5461 ggttcccggc tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag
     5521 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5581 ggcagcgccg tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc
     5641 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5701 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5761 taggctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     5821 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     5881 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     5941 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     6001 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     6061 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     6121 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6181 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     6241 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     6301 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6361 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     6421 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6481 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     6541 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6601 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6661 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     6721 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6781 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6841 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6901 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6961 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     7021 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     7081 tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac tctggctcta
     7141 tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc
     7201 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
     7261 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
     7321 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     7381 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
     7441 tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta
     7501 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     7561 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     7621 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7681 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     7741 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     7801 atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg
     7861 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     7921 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     7981 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
     8041 gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg
     8101 ccgc
//
