LOCUS       sequence-295997-        5151 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 5151)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5151)
  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-m4133547
FEATURES             Location/Qualifiers
     source          1..5151
                     /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             923..967
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1020..1141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1031..1050)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1085..1104
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1256..1302
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1332..1468
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1513..1584
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1810..2398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1890..1909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3144..3212,3213..4004)
                     /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(3362..3381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4017..4153
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4305..4376
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4381..4427)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4445..4493
                     /note="synthetic polyadenylation signal"
     enhancer        4574..4877
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4879..5081
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5032..5052
                     /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 cgaaagaaac cgcggcagct aagttcgaga ggcagcacat
      961 ggacagttaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1021 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1081 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1141 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1201 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1261 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1321 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1381 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1441 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1501 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1561 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1621 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1681 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1741 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1801 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1861 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1921 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1981 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2041 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2101 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2161 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2221 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2281 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2341 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2401 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2461 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2521 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2581 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2641 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2701 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2761 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2821 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2881 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     2941 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     3001 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     3061 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     3121 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     3181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3241 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     3301 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     3361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3421 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     3481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3541 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     3601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     3721 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     3781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     4021 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     4081 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     4141 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     4201 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     4261 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     4321 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     4381 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     4441 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     4501 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     4561 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4621 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4681 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4741 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4801 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4861 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4921 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     4981 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5041 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     5101 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
