LOCUS       sequence-295034-        6189 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 6189)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6189)
  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-m4134143
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1916..1936
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1940..2005
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2058..2179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2069..2088)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2123..2142
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2294..2340
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2370..2506
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2551..2622
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2848..3436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2928..2947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4182..4250,4251..5042)
                     /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(4400..4419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5055..5191
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5343..5414
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5419..5465)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5483..5531
                     /note="synthetic polyadenylation signal"
     enhancer        5612..5915
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5917..6119
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6070..6090
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac actcgctatg tcgataacaa cttctgtggc cctgatggct accctcttga
       61 gtgcattaaa gaccttctag cacgtgctgg taaagcttca tgcactttgt ccgaacaact
      121 ggactttatt gacactaaga ggggtgtata ctgctgccgt gaacatgagc atgaaattgc
      181 ttggtacacg gaacgttctg aaaagagcta tgaattgcag acaccttttg aaattaaatt
      241 ggcaaagaaa tttgacacct tcaatgggga atgtccaaat tttgtatttc ccttaaattc
      301 cataatcaag actattcaac caagggttga aaagaaaaag cttgatggct ttatgggtag
      361 aattcgatct gtctatccag ttgcgtcacc aaatgaatgc aaccaaatgt gcctttcaac
      421 tctcatgaag tgtgatcatt gtggtgaaac ttcatggcag acgggcgatt ttgttaaagc
      481 cacttgcgaa ttttgtggca ctgagaattt gactaaagaa ggtgccacta cttgtggtta
      541 cttaccccaa aatgctgttg ttaaaattta ttgtccagca tgtcacaatt cagaagtagg
      601 acctgagcat agtcttgccg aataccataa tgaatctggc ttgaaaacca ttcttcgtaa
      661 gggtggtcgc actattgcct ttggaggctg tgtgttctct tatgttggtt gccataacaa
      721 gtgtgcctat tgggttccac gtgctagcgc taacataggt tgtaaccata caggtgttgt
      781 tggagaaggt tccgaaggtc ttaatgacaa ccttcttgaa atactccaaa aagagaaagt
      841 caacatcaat attgttggtg actttaaact taatgaagag atcgccatta ttttggcatc
      901 tttttctgct tccacaagtg cttttgtgga aactgtgaaa ggtttggatt ataaagcatt
      961 caaacaaatt gttgaatcct gtggtaattt taaagttaca aaaggaaaag ctaaaaaagg
     1021 tgcctggaat attggtgaac agaaatcaat actgagtcct ctttatgcat ttgcatcaga
     1081 ggctgctcgt gttgtacgat caattttctc ccgcactctt gaaactgctc aaaattctgt
     1141 gcgtgtttta cagaaggccg ctataacaat actagatgga atttcacagt attcactgag
     1201 actcattgat gctatgatgt tcacatctga tttggctact aacaatctag ttgtaatggc
     1261 ctacattaca ggtggtgttg ttcagttgac ttcgcagtgg ctaactaaca tctttggcac
     1321 tgtttatgaa aaactcaaac ccgtccttga ttggcttgaa gagaagttta aggaaggtgt
     1381 agagtttctt agagacggtt gggaaattgt taaatttatc tcaacctgtg cttgtgaaat
     1441 tgtcggtgga caaattgtca cctgtgcaaa ggaaattaag gagagtgttc agacattctt
     1501 taagcttgta aataaatttt tggctttgtg tgctgactct atcattattg gtggagctaa
     1561 acttaaagcc ttgaatttag gtgaaacatt tgtcacgcac tcaaagggat tgtacagaaa
     1621 gtgtgttaaa tccagagaag aaactggcct actcatgcct ctaaaagccc caaaagaaat
     1681 tatcttctta gagggagaaa cacttcccac agaagtgtta acagaggaag ttgtcttgaa
     1741 aactggtgat ttacaaccat tagaacaacc tactagtgaa gctgttgaag ctccattggt
     1801 tggtacacca gtttgtatta acgggcttat gttgctcgaa atcaaagaca cagaaaagta
     1861 ctgtgccctt gcacctaata tgatggtaac aaacaatacc ttcacactca aaggcgaaaa
     1921 cctttacttt cagtcgatgg attataagga ccacgatgga gactacaaag atcatgacat
     1981 tgattacaag gacgacgatg acaaataagc tagcttgact gactgagata cagcgtacct
     2041 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2101 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2161 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2221 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
     2281 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2341 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
     2401 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     2461 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
     2521 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     2581 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     2641 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     2701 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     2761 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     2821 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2881 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2941 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3001 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3061 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3121 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3181 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3241 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3301 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3361 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3421 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3481 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3541 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3601 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3661 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     3721 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     3781 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     3841 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     3901 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag
     3961 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     4021 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     4081 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     4141 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     4201 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4261 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     4321 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     4381 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4441 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     4501 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4561 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     4621 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4681 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4741 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     4801 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4861 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4921 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4981 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5041 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     5101 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     5161 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     5221 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     5281 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     5341 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     5401 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     5461 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     5521 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     5581 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     5641 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     5701 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     5761 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     5821 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     5881 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     5941 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     6001 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     6061 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     6121 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     6181 agcggccgc
//
