LOCUS       sequence-307276-        7521 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 7521)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7521)
  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-m4133211
FEATURES             Location/Qualifiers
     source          1..7521
                     /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             920..985
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1054..1175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1065..1084)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1119..1138
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1290..1336
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1366..1502
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1547..1618
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1844..2432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1924..1943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2932..4141
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3978..3997
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4145..5173
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5200..5248
                     /note="synthetic polyadenylation signal"
     CDS             join(5514..5582,5583..6374)
                     /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(5732..5751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6387..6523
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6675..6746
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6751..6797)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6815..6863
                     /note="synthetic polyadenylation signal"
     enhancer        6944..7247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7249..7451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7402..7422
                     /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 cagtcgatgg attataagga ccacgatgga gactacaaag atcatgacat
      961 tgattacaag gacgacgatg acaaataagg atccgcaggc ctctgctagc ttgactgact
     1021 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
     1081 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1141 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1201 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1261 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     1321 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     1381 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     1441 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     1501 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     1561 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     1621 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     1681 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     1741 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     1801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2401 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2461 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2641 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     2701 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     2761 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     2821 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     2881 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     2941 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     3001 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     3061 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     3121 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     3181 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     3241 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3301 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     3361 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     3421 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     3481 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     3541 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     3601 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     3661 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     3721 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     3781 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     3841 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     3901 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     3961 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     4021 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     4081 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     4141 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4201 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4261 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4321 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     4381 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4441 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4501 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4561 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4621 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4681 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4741 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4801 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4861 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4921 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4981 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5041 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     5101 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5161 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     5221 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     5281 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     5341 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     5401 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     5461 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     5521 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5581 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     5641 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5701 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5761 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5821 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5881 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     5941 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6001 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6061 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     6121 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6181 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6241 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6301 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6361 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     6421 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     6481 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     6541 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     6601 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     6661 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     6721 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     6781 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     6841 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     6901 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     6961 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7021 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     7081 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     7141 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     7201 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     7261 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     7321 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     7381 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     7441 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     7501 gacacacccg ccagcggccg c
//
