LOCUS       sequence-295613-        8575 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 8575)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8575)
  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-m4134367
FEATURES             Location/Qualifiers
     source          1..8575
                     /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"
     promoter        4422..5631
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5468..5487
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5635..6234
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5635..5654)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6131..6151
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6261..6309
                     /note="synthetic polyadenylation signal"
     CDS             join(6575..6643,6644..7435)
                     /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(6793..6812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7448..7584
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7729..7800
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7805..7851)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7869..7917
                     /note="synthetic polyadenylation signal"
     enhancer        7998..8301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8303..8505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8456..8476
                     /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 tggcctccgc gccgggtttt
     4441 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     4501 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     4561 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     4621 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     4681 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     4741 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     4801 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     4861 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     4921 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     4981 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     5041 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     5101 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     5161 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     5221 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     5281 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     5341 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     5401 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     5461 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     5521 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     5581 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     5641 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5701 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5761 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5821 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5881 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5941 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     6001 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     6061 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     6121 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     6181 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     6241 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     6301 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     6361 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     6421 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     6481 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     6541 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     6601 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6661 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     6721 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     6781 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6841 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     6901 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6961 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     7021 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7081 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     7141 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     7201 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7261 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7321 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7381 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     7441 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     7501 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     7561 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     7621 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     7681 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     7741 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     7801 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     7861 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     7921 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     7981 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     8041 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     8101 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8161 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8221 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     8281 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     8341 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     8401 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     8461 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     8521 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
