LOCUS       sequence-295963-        9020 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 9020)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9020)
  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-m4134355
FEATURES             Location/Qualifiers
     source          1..9020
                     /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    2560..2681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2571..2590)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2625..2644
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2796..2842
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2872..3008
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3053..3124
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3350..3938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3430..3449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4438..5647
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5484..5503
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5651..6679
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6706..6754
                     /note="synthetic polyadenylation signal"
     CDS             join(7020..7088,7089..7880)
                     /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(7238..7257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7893..8029
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8174..8245
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8250..8296)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8314..8362
                     /note="synthetic polyadenylation signal"
     enhancer        8443..8746
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8748..8950
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8901..8921
                     /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 ataaggatcc gcaggcctct gctagcttga
     2521 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
     2581 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2641 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2701 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     2761 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
     2821 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
     2881 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     2941 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     3001 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     3061 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     3121 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     3181 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     3241 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     3301 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3361 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3421 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3481 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3541 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3601 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3661 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3721 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3781 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3841 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3901 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3961 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4021 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4081 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4141 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     4201 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     4261 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     4321 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     4381 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc
     4441 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca
     4501 cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     4561 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     4621 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     4681 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     4741 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     4801 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     4861 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     4921 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     4981 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     5041 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct
     5101 aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt
     5161 gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg
     5221 cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac
     5281 gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta
     5341 ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca
     5401 ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt
     5461 atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc
     5521 gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg
     5581 taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt
     5641 gttagacaac atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     5701 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     5761 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     5821 tttctacaaa gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga
     5881 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     5941 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     6001 ggaggcaatg gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt
     6061 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     6121 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     6181 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     6241 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     6301 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     6361 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     6421 tgcaggatcg ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     6481 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     6541 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt
     6601 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     6661 tccgagggca aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc
     6721 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     6781 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     6841 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     6901 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     6961 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     7021 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     7081 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     7141 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     7201 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     7261 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7321 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7381 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     7441 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7501 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7561 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     7621 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7681 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7741 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7801 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7861 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     7921 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     7981 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcgtttaaa
     8041 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     8101 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     8161 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     8221 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     8281 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     8341 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
     8401 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     8461 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     8521 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     8581 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     8641 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     8701 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
     8761 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     8821 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     8881 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     8941 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
     9001 acacacccgc cagcggccgc
//
