LOCUS       sequence-295268-        5145 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 5145)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5145)
  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-m4134146
FEATURES             Location/Qualifiers
     source          1..5145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             896..916
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             917..961
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1014..1135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1025..1044)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1079..1098
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1250..1296
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1326..1462
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1507..1578
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1804..2392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1884..1903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3138..3206,3207..3998)
                     /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(3356..3375)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4011..4147
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4299..4370
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4375..4421)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4439..4487
                     /note="synthetic polyadenylation signal"
     enhancer        4568..4871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4873..5075
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5026..5046
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt caagcgtggc aaccgggtgt tgctatgcct aatctttaca aaatgcaaag
       61 aatgctatta gaaaagtgtg accttcaaaa ttatggtgat agtgcaacat tacctaaagg
      121 cataatgatg aatgtcgcaa aatatactca actgtgtcaa tatttaaaca cattaacatt
      181 agctgtaccc tataatatga gagttataca ttttggtgct ggttctgata aaggagttgc
      241 accaggtaca gctgttttaa gacagtggtt gcctacgggt acgctgcttg tcgattcaga
      301 tcttaatgac tttgtctctg atgcagattc aactttgatt ggtgattgtg caactgtaca
      361 tacagctaat aaatgggatc tcattattag tgatatgtac gaccctaaga ctaaaaatgt
      421 tacaaaagaa aatgactcta aagagggttt tttcacttac atttgtgggt ttatacaaca
      481 aaagctagct cttggaggtt ccgtggctat aaagataaca gaacattctt ggaatgctga
      541 tctttataag ctcatgggac acttcgcatg gtggacagcc tttgttacta atgtgaatgc
      601 gtcatcatct gaagcatttt taattggatg taattatctt ggcaaaccac gcgaacaaat
      661 agatggttat gtcatgcatg caaattacat attttggagg aatacaaatc caattcagtt
      721 gtcttcctat tctttatttg acatgagtaa atttcccctt aaattaaggg gtactgctgt
      781 tatgtcttta aaagaaggtc aaatcaatga tatgatttta tctcttctta gtaaaggtag
      841 acttataatt agagaaaaca acagagttgt tatttctagt gatgttcttg ttaacgaaaa
      901 cctttacttt cagtcgaaag aaaccgcggc agctaagttc gagaggcagc acatggacag
      961 ttaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1021 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1081 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1141 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1201 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     1261 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1321 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     1381 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     1441 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     1501 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     1561 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     1621 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     1681 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     1741 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     2641 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     2701 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     2761 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     2821 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     2881 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     2941 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     3001 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     3061 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     3121 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3181 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3241 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3301 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     3361 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3421 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     3481 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3541 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3601 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3661 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     3721 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     3781 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3841 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3901 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3961 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     4021 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     4081 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     4141 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     4201 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     4261 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     4321 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     4381 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     4441 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     4501 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4561 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4621 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4681 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4741 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4801 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4861 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4921 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4981 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5041 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     5101 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
