LOCUS       sequence-319571-        8054 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 8054)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8054)
  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-m4133398
FEATURES             Location/Qualifiers
     source          1..8054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2189..2209
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2210..2236
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2289..2410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2300..2319)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2354..2373
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2525..2571
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2601..2737
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2782..2853
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3079..3667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3159..3178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4167..5376
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5213..5232
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5380..5979
                     /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(5380..5399)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5876..5896
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6006..6054
                     /note="synthetic polyadenylation signal"
     CDS             join(6320..6388,6389..7180)
                     /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(6538..6557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7193..7329
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7481..7552
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7557..7603)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7621..7669
                     /note="synthetic polyadenylation signal"
     enhancer        7750..8053
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
      241 taccatccac tcgacacacc cgccagcggc cgccaccatg tacactcgct atgtcgataa
      301 caacttctgt ggccctgatg gctaccctct tgagtgcatt aaagaccttc tagcacgtgc
      361 tggtaaagct tcatgcactt tgtccgaaca actggacttt attgacacta agaggggtgt
      421 atactgctgc cgtgaacatg agcatgaaat tgcttggtac acggaacgtt ctgaaaagag
      481 ctatgaattg cagacacctt ttgaaattaa attggcaaag aaatttgaca ccttcaatgg
      541 ggaatgtcca aattttgtat ttcccttaaa ttccataatc aagactattc aaccaagggt
      601 tgaaaagaaa aagcttgatg gctttatggg tagaattcga tctgtctatc cagttgcgtc
      661 accaaatgaa tgcaaccaaa tgtgcctttc aactctcatg aagtgtgatc attgtggtga
      721 aacttcatgg cagacgggcg attttgttaa agccacttgc gaattttgtg gcactgagaa
      781 tttgactaaa gaaggtgcca ctacttgtgg ttacttaccc caaaatgctg ttgttaaaat
      841 ttattgtcca gcatgtcaca attcagaagt aggacctgag catagtcttg ccgaatacca
      901 taatgaatct ggcttgaaaa ccattcttcg taagggtggt cgcactattg cctttggagg
      961 ctgtgtgttc tcttatgttg gttgccataa caagtgtgcc tattgggttc cacgtgctag
     1021 cgctaacata ggttgtaacc atacaggtgt tgttggagaa ggttccgaag gtcttaatga
     1081 caaccttctt gaaatactcc aaaaagagaa agtcaacatc aatattgttg gtgactttaa
     1141 acttaatgaa gagatcgcca ttattttggc atctttttct gcttccacaa gtgcttttgt
     1201 ggaaactgtg aaaggtttgg attataaagc attcaaacaa attgttgaat cctgtggtaa
     1261 ttttaaagtt acaaaaggaa aagctaaaaa aggtgcctgg aatattggtg aacagaaatc
     1321 aatactgagt cctctttatg catttgcatc agaggctgct cgtgttgtac gatcaatttt
     1381 ctcccgcact cttgaaactg ctcaaaattc tgtgcgtgtt ttacagaagg ccgctataac
     1441 aatactagat ggaatttcac agtattcact gagactcatt gatgctatga tgttcacatc
     1501 tgatttggct actaacaatc tagttgtaat ggcctacatt acaggtggtg ttgttcagtt
     1561 gacttcgcag tggctaacta acatctttgg cactgtttat gaaaaactca aacccgtcct
     1621 tgattggctt gaagagaagt ttaaggaagg tgtagagttt cttagagacg gttgggaaat
     1681 tgttaaattt atctcaacct gtgcttgtga aattgtcggt ggacaaattg tcacctgtgc
     1741 aaaggaaatt aaggagagtg ttcagacatt ctttaagctt gtaaataaat ttttggcttt
     1801 gtgtgctgac tctatcatta ttggtggagc taaacttaaa gccttgaatt taggtgaaac
     1861 atttgtcacg cactcaaagg gattgtacag aaagtgtgtt aaatccagag aagaaactgg
     1921 cctactcatg cctctaaaag ccccaaaaga aattatcttc ttagagggag aaacacttcc
     1981 cacagaagtg ttaacagagg aagttgtctt gaaaactggt gatttacaac cattagaaca
     2041 acctactagt gaagctgttg aagctccatt ggttggtaca ccagtttgta ttaacgggct
     2101 tatgttgctc gaaatcaaag acacagaaaa gtactgtgcc cttgcaccta atatgatggt
     2161 aacaaacaat accttcacac tcaaaggcga aaacctttac tttcagtcgt acccctatga
     2221 cgtaccagat tatgcgtaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
     2281 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2341 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2401 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2461 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     2521 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     2581 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     2641 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     2701 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     2761 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     2821 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     2881 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     2941 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     3001 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3901 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     3961 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     4021 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     4081 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     4141 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     4201 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     4261 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     4321 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     4381 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     4441 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     4501 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     4561 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     4621 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     4681 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     4741 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     4801 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     4861 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     4921 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     4981 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     5041 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     5101 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     5161 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     5221 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     5281 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     5341 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     5401 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     5461 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     5521 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5581 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5641 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5701 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5761 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5821 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5881 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5941 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     6001 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     6061 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     6121 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     6181 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     6241 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     6301 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     6361 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6421 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6481 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     6541 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6601 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     6661 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6721 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6781 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6841 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6901 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     6961 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7021 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7081 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7141 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     7201 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     7261 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     7321 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     7381 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     7441 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     7501 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     7561 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     7621 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     7681 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     7741 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     7801 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7861 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7921 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     7981 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     8041 cgctattacc atgc
//
