LOCUS       sequence-295993-        5661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5661)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5661)
  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-m4133481
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             902..922
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             956..1033
                     /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             1061..1081
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1530..1651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1541..1560)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1595..1614
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1766..1812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1842..1978
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2023..2094
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2320..2908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2400..2419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3654..3722,3723..4514)
                     /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(3872..3891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4527..4663
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4815..4886
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4891..4937)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4955..5003
                     /note="synthetic polyadenylation signal"
     enhancer        5084..5387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5389..5591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5542..5562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtcc tcacaagcgt ggcagccagg cgtagcaatg cctaatttat acaagatgca
       61 gcggatgctc cttgaaaaat gcgacctaca gaactatgga gatagtgcta cattgcccaa
      121 ggggataatg atgaatgtgg ccaaatatac ccaactgtgt cagtacctga acactctcac
      181 gctggccgtt ccgtacaaca tgagggtgat tcattttggt gccggcagcg acaagggagt
      241 cgctcccggg accgctgtgc tgcgacagtg gcttcctaca ggcactctgc tagtggattc
      301 tgatttgaat gacttcgtca gcgacgcaga ctcgaccctc atcggagatt gtgcgactgt
      361 tcacacagcc aacaaatggg acctgatcat ttccgatatg tatgatccaa agaccaaaaa
      421 tgtcacgaag gagaacgact caaaggaggg tttctttacg tacatctgcg ggttcatcca
      481 acagaaactt gcactgggcg gaagtgtggc aattaaaata acagaacact cttggaacgc
      541 cgacctatac aagcttatgg gtcattttgc ttggtggacc gcctttgtta ctaatgtaaa
      601 cgcttcaagc tccgaggcct tcctgatagg ttgtaattat ttaggcaagc ctagagagca
      661 gattgatggg tatgtgatgc acgcaaatta catcttctgg cgcaacacaa atccaattca
      721 gctcagttct tattcactgt ttgatatgag caaattcccc cttaaactga gaggcactgc
      781 cgtcatgtcc ttgaaggaag gacaaatcaa cgatatgatc ttgtctctcc ttagcaaggg
      841 gcgtttgatt attagggaga acaatcgggt ggtgatctca tcggacgttc tggtcaacaa
      901 tgaaaacctt tactttcagt cgggacggcg aatcccgggg ctgataaatc cctggaaaag
      961 gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc
     1021 aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct atttccaggg
     1081 tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata aagaacagca
     1141 gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac gcaacgcttt
     1201 cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg aagccaagaa
     1261 gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc agcagaatgc
     1321 tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg catttatcca
     1381 gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca aaaagctcaa
     1441 tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga ctgactgaga
     1501 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1561 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1621 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1681 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1741 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1801 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1861 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1921 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1981 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     2041 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     2101 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     2161 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     2221 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     2281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3121 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     3181 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     3241 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     3301 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3361 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     3421 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     3481 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     3541 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3601 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     3661 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3721 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3781 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3841 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3901 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3961 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4021 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     4081 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4141 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4201 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     4261 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4321 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4381 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4441 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4501 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     4561 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4621 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     4681 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4741 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     4801 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     4861 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     4921 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     4981 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     5041 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     5101 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     5161 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     5221 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     5281 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     5341 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     5401 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     5461 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     5521 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     5581 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     5641 gacacacccg ccagcggccg c
//
