LOCUS       sequence-307283-        6510 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 6510)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6510)
  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-m4133357
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      172..308
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    460..531
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(536..582)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    600..648
                     /note="synthetic polyadenylation signal"
     enhancer        729..1032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1034..1236
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1187..1207
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3222..3242
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3591..3608
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3696..3715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3750..3769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3921..3967
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3997..4133
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4178..4249
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4475..5063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4555..4574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5809..5877,5878..6510,1..159)
                     /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(6027..6046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
       61 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
      121 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
      181 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
      241 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
      301 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
      361 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
      421 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
      481 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
      541 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
      601 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
      661 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
      721 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      781 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      841 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      901 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      961 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1021 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     1081 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1141 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1201 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     1261 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc atgtacactc
     1321 gctatgtcga taacaacttc tgtggccctg atggctaccc tcttgagtgc attaaagacc
     1381 ttctagcacg tgctggtaaa gcttcatgca ctttgtccga acaactggac tttattgaca
     1441 ctaagagggg tgtatactgc tgccgtgaac atgagcatga aattgcttgg tacacggaac
     1501 gttctgaaaa gagctatgaa ttgcagacac cttttgaaat taaattggca aagaaatttg
     1561 acaccttcaa tggggaatgt ccaaattttg tatttccctt aaattccata atcaagacta
     1621 ttcaaccaag ggttgaaaag aaaaagcttg atggctttat gggtagaatt cgatctgtct
     1681 atccagttgc gtcaccaaat gaatgcaacc aaatgtgcct ttcaactctc atgaagtgtg
     1741 atcattgtgg tgaaacttca tggcagacgg gcgattttgt taaagccact tgcgaatttt
     1801 gtggcactga gaatttgact aaagaaggtg ccactacttg tggttactta ccccaaaatg
     1861 ctgttgttaa aatttattgt ccagcatgtc acaattcaga agtaggacct gagcatagtc
     1921 ttgccgaata ccataatgaa tctggcttga aaaccattct tcgtaagggt ggtcgcacta
     1981 ttgcctttgg aggctgtgtg ttctcttatg ttggttgcca taacaagtgt gcctattggg
     2041 ttccacgtgc tagcgctaac ataggttgta accatacagg tgttgttgga gaaggttccg
     2101 aaggtcttaa tgacaacctt cttgaaatac tccaaaaaga gaaagtcaac atcaatattg
     2161 ttggtgactt taaacttaat gaagagatcg ccattatttt ggcatctttt tctgcttcca
     2221 caagtgcttt tgtggaaact gtgaaaggtt tggattataa agcattcaaa caaattgttg
     2281 aatcctgtgg taattttaaa gttacaaaag gaaaagctaa aaaaggtgcc tggaatattg
     2341 gtgaacagaa atcaatactg agtcctcttt atgcatttgc atcagaggct gctcgtgttg
     2401 tacgatcaat tttctcccgc actcttgaaa ctgctcaaaa ttctgtgcgt gttttacaga
     2461 aggccgctat aacaatacta gatggaattt cacagtattc actgagactc attgatgcta
     2521 tgatgttcac atctgatttg gctactaaca atctagttgt aatggcctac attacaggtg
     2581 gtgttgttca gttgacttcg cagtggctaa ctaacatctt tggcactgtt tatgaaaaac
     2641 tcaaacccgt ccttgattgg cttgaagaga agtttaagga aggtgtagag tttcttagag
     2701 acggttggga aattgttaaa tttatctcaa cctgtgcttg tgaaattgtc ggtggacaaa
     2761 ttgtcacctg tgcaaaggaa attaaggaga gtgttcagac attctttaag cttgtaaata
     2821 aatttttggc tttgtgtgct gactctatca ttattggtgg agctaaactt aaagccttga
     2881 atttaggtga aacatttgtc acgcactcaa agggattgta cagaaagtgt gttaaatcca
     2941 gagaagaaac tggcctactc atgcctctaa aagccccaaa agaaattatc ttcttagagg
     3001 gagaaacact tcccacagaa gtgttaacag aggaagttgt cttgaaaact ggtgatttac
     3061 aaccattaga acaacctact agtgaagctg ttgaagctcc attggttggt acaccagttt
     3121 gtattaacgg gcttatgttg ctcgaaatca aagacacaga aaagtactgt gcccttgcac
     3181 ctaatatgat ggtaacaaac aataccttca cactcaaagg cgaaaacctt tactttcagt
     3241 cgatgagtga caaaatcatt catctaaccg atgactcctt tgatacagac gtgttgaagg
     3301 ccgatggtgc gatactggta gatttctggg ctgaatggtg tggcccgtgc aagatgatcg
     3361 cacccattct ggacgagatc gccgacgagt atcaaggaaa attaactgtc gctaagctta
     3421 atatagatca gaacccaggg acggcaccta aatacggcat caggggaata cctaccctcc
     3481 tgctctttaa gaatggggaa gttgccgcga caaaagtggg agccctgtct aagggccagc
     3541 taaaagagtt cttggacgct aacctggcag gttcaggctc ggggcacatg caccatcacc
     3601 atcaccatag cagcgggctc gtgccacgcg gctaagctag cttgactgac tgagatacag
     3661 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     3721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3781 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3841 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     3901 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3961 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     4021 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     4081 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     4141 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     4201 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     4261 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     4321 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     4381 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     4441 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4501 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4561 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4621 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4681 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4741 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4801 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4861 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4921 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4981 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5041 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5101 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5161 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5221 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5281 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     5341 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     5401 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     5461 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     5521 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc
     5581 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     5641 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     5701 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     5761 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     5821 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5881 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     5941 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     6001 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6061 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     6121 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6181 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     6241 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6301 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6361 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     6421 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6481 gctggctggt ttattgctga taaatctgga
//
