LOCUS       sequence-295199-        6675 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 6675)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6675)
  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-m4133702
FEATURES             Location/Qualifiers
     source          1..6675
                     /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             1970..2047
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2075..2095
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2544..2665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2555..2574)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2609..2628
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2780..2826
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2856..2992
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3037..3108
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3334..3922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3414..3433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4668..4736,4737..5528)
                     /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(4886..4905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5541..5677
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5829..5900
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5905..5951)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5969..6017
                     /note="synthetic polyadenylation signal"
     enhancer        6098..6401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6403..6605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6556..6576
                     /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 cagtcgggac ggcgaatccc ggggctgata aatccctgga aaaggagatg
     1981 gaagaagaac tttattgcag tgtctgcggc taaccggttc aaaaagatca gttcaagcgg
     2041 cgccctagat tacgacattc ctacgaccgc ctccgagaat ctctatttcc agggtgaatt
     2101 tggcttggca caacatgacg aggccgtcga taacaagttt aataaagaac agcagaacgc
     2161 tttctatgag atccttcact taccaaatct gaacgaagag caacgcaacg ctttcattca
     2221 gagcctgaaa gacgatccct cgcagtctgc aaatctcctg gccgaagcca agaagctgaa
     2281 tgacgcacag gctccaaaag ttgataacaa gtttaacaag gagcagcaga atgctttcta
     2341 cgagatattg caccttccta atctgaatga ggaacaaaga aatgcattta tccagtccct
     2401 aaaagacgat ccatcacaga gtgccaacct gcttgcggaa gccaaaaagc tcaatgatgc
     2461 ccaggccccg aaagtagacg cgaaccatca ataagctagc ttgactgact gagatacagc
     2521 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2581 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2641 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2701 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     2761 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     2821 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     2881 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     2941 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     3001 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     3061 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     3121 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     3181 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     3241 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     3301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3961 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4021 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4081 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4141 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     4201 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     4261 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     4321 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     4381 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     4441 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4501 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4561 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4621 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     4861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     4981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5041 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     5101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5221 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5281 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5521 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5581 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5641 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5701 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5761 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     5821 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     5881 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     5941 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     6001 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     6061 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6361 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     6421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6601 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6661 cccgccagcg gccgc
//
