LOCUS       sequence-295919-        7478 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 7478)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7478)
  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-m4133490
FEATURES             Location/Qualifiers
     source          1..7478
                     /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..949
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1018..1139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1029..1048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1083..1102
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1254..1300
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1330..1466
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1511..1582
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1808..2396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1888..1907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2896..4105
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3942..3961
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4109..5137
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5164..5212
                     /note="synthetic polyadenylation signal"
     CDS             join(5478..5546,5547..6338)
                     /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(5696..5715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6351..6487
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6632..6703
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6708..6754)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6772..6820
                     /note="synthetic polyadenylation signal"
     enhancer        6901..7204
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7206..7408
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7359..7379
                     /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 cgtaccccta tgacgtacca gattatgcgt aaggatccgc
      961 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
     1021 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1081 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1141 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1201 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
     1261 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
     1321 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     1381 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     1441 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
     1501 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
     1561 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
     1621 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     1681 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     1741 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1801 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1861 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1921 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1981 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2041 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2101 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2161 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2221 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2281 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2341 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2401 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2461 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2521 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2581 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     2641 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     2701 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     2761 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     2821 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2881 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2941 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     3001 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     3061 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     3121 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     3181 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     3241 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     3301 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     3361 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     3421 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     3481 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     3541 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     3601 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     3661 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     3721 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     3781 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3841 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3901 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3961 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     4021 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     4081 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     4141 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     4201 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     4261 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     4321 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     4381 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     4441 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     4501 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     4561 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4621 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4681 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4741 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4801 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4861 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4921 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4981 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     5041 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     5101 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     5161 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     5221 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     5281 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     5341 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     5401 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     5461 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5521 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5581 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5641 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5701 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5761 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5821 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5881 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5941 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6001 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     6061 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     6121 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6181 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6241 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6301 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     6361 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     6421 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     6481 tccttaagat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     6541 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     6601 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     6661 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     6721 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     6781 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     6841 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6901 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6961 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     7021 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     7081 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     7141 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     7201 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     7261 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     7321 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     7381 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     7441 gatcctacca tccactcgac acacccgcca gcggccgc
//
