LOCUS       sequence-295662-        6195 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 6195)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6195)
  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-m4134528
FEATURES             Location/Qualifiers
     source          1..6195
                     /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"
     CDS             join(4188..4256,4257..5048)
                     /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(4406..4425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5061..5197
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5349..5420
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5425..5471)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5489..5537
                     /note="synthetic polyadenylation signal"
     enhancer        5618..5921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5923..6125
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6076..6096
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6194..6195,1..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 tggtgtaccg agaacgatcc
     3961 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4021 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4081 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4141 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4201 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4261 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4321 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     4381 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4441 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     4501 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4561 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     4621 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4681 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4741 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     4801 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4861 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4921 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4981 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5041 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5101 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5161 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5221 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5281 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     5341 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     5401 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     5461 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     5521 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     5581 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     5641 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     5701 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     5761 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     5821 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     5881 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     5941 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6001 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6061 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6121 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6181 cccgccagcg gccgc
//
