LOCUS       sequence-295516-        5496 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 5496)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5496)
  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-m4133778
FEATURES             Location/Qualifiers
     source          1..5496
                     /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             1271..1288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1365..1486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1376..1395)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1430..1449
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1601..1647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1677..1813
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1858..1929
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2155..2743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2235..2254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3489..3557,3558..4349)
                     /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(3707..3726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4362..4498
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4650..4721
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4726..4772)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4790..4838
                     /note="synthetic polyadenylation signal"
     enhancer        4919..5222
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5224..5426
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5377..5397
                     /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 cgatgagtga caaaatcatt catctaaccg atgactcctt
      961 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
     1021 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
     1081 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
     1141 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
     1201 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
     1261 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaagctag
     1321 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     1381 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1441 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1501 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1561 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1621 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     1681 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1741 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1801 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1861 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1921 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1981 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     2041 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     2101 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2161 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2221 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2281 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2341 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2401 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2461 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2521 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2581 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2641 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2701 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2761 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2821 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2881 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2941 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     3001 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     3061 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     3121 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     3181 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     3241 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     3301 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     3361 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     3421 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     3481 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3601 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3661 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3721 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3781 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3901 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4021 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     4081 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     4141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4321 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     4381 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     4441 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     4501 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     4561 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     4621 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     4681 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     4741 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     4801 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     4861 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4921 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4981 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     5041 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     5101 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     5161 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     5221 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     5281 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5341 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5401 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     5461 tcctaccatc cactcgacac acccgccagc ggccgc
//
