LOCUS       sequence-295647-        8095 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 8095)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8095)
  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-m4134392
FEATURES             Location/Qualifiers
     source          1..8095
                     /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             1946..2011
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2064..2185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2075..2094)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2129..2148
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2300..2346
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2376..2512
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2557..2628
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2854..3442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2934..2953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3942..5151
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4988..5007
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5155..5754
                     /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(5155..5174)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5651..5671
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5781..5829
                     /note="synthetic polyadenylation signal"
     CDS             join(6095..6163,6164..6955)
                     /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(6313..6332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6968..7104
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7249..7320
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7325..7371)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7389..7437
                     /note="synthetic polyadenylation signal"
     enhancer        7518..7821
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7823..8025
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7976..7996
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8094..8095,1..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 cgatggatta taaggaccac gatggagact acaaagatca
     1981 tgacattgat tacaaggacg acgatgacaa ataagctagc ttgactgact gagatacagc
     2041 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2101 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2161 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2221 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     2281 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     2341 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     2401 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     2461 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     2521 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     2581 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     2641 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     2701 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     2761 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     2821 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2881 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2941 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3001 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3061 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3121 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3181 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3241 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3301 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3361 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3421 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3481 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3541 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3601 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3661 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     3721 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     3781 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3841 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3901 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     3961 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     4021 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     4081 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     4141 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     4201 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     4261 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     4321 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     4381 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     4441 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     4501 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     4561 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     4621 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     4681 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     4741 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     4801 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     4861 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     4921 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     4981 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     5041 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     5101 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     5161 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5221 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5281 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5341 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5401 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5461 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     5521 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5581 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     5641 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     5701 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     5761 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     5821 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     5881 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     5941 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     6001 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     6061 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     6121 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6181 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     6241 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     6301 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6361 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     6421 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6481 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     6541 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6601 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     6661 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     6721 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6781 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6841 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6901 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     6961 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     7021 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     7081 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     7141 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     7201 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     7261 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     7321 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     7381 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     7441 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     7501 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     7561 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     7621 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     7681 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     7741 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     7801 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     7861 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     7921 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     7981 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     8041 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
