LOCUS       sequence-295880-        6198 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 6198)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6198)
  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-m4134380
FEATURES             Location/Qualifiers
     source          1..6198
                     /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             1937..2014
                     /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"
     polyA_signal    2067..2188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2078..2097)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2132..2151
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2303..2349
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2379..2515
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2560..2631
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2857..3445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2937..2956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4191..4259,4260..5051)
                     /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(4409..4428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5064..5200
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5352..5423
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5428..5474)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5492..5540
                     /note="synthetic polyadenylation signal"
     enhancer        5621..5924
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5926..6128
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6079..6099
                     /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 cagtcgaaga ggcgatggaa aaagaatttt atcgcggtga gtgcagccaa
     1981 cagattcaaa aagatatcct cgtctggtgc cctctaagct agcttgactg actgagatac
     2041 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
     2101 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2161 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2221 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     2281 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     2341 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     2401 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     2461 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     2521 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     2581 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     2641 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     2701 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     2761 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     2821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3421 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3541 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3601 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3661 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     3721 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     3781 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     3841 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     3901 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     3961 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     4021 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     4081 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     4141 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     4201 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4261 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     4321 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     4381 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4441 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     4501 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4561 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     4621 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4681 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4741 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4801 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4861 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4921 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4981 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5041 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     5101 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     5161 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     5221 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     5281 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     5341 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     5401 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     5461 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     5521 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     5581 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     5641 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     5701 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     5761 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     5821 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     5881 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     5941 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     6001 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     6061 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     6121 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     6181 acacccgcca gcggccgc
//
