LOCUS       sequence-295636-        8581 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 8581)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8581)
  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-m4134423
FEATURES             Location/Qualifiers
     source          1..8581
                     /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"
     promoter        4428..5637
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5474..5493
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5641..6240
                     /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(5641..5660)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6137..6157
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6267..6315
                     /note="synthetic polyadenylation signal"
     CDS             join(6581..6649,6650..7441)
                     /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(6799..6818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7454..7590
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7735..7806
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7811..7857)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7875..7923
                     /note="synthetic polyadenylation signal"
     enhancer        8004..8307
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8309..8511
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8462..8482
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8580..8581,1..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 cgcgcctggc ctccgcgccg
     4441 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     4501 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     4561 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     4621 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     4681 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     4741 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     4801 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     4861 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     4921 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     4981 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     5041 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     5101 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     5161 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     5221 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     5281 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     5341 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     5401 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     5461 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     5521 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     5581 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     5641 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     5701 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5761 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5821 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5881 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5941 tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag
     6001 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     6061 agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg
     6121 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     6181 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag
     6241 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     6301 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     6361 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     6421 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     6481 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     6541 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     6601 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6661 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     6721 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     6781 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6841 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     6901 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6961 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     7021 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7081 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7141 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     7201 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7261 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7321 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7381 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7441 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     7501 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     7561 gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct ggctctatcg
     7621 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     7681 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     7741 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     7801 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     7861 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     7921 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     7981 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     8041 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     8101 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     8161 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     8221 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     8281 tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt
     8341 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     8401 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     8461 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     8521 aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg
     8581 c
//
