LOCUS       sequence-295991-        7469 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 7469)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7469)
  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-m4133310
FEATURES             Location/Qualifiers
     source          1..7469
                     /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..940
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1009..1130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1020..1039)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1074..1093
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1245..1291
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1321..1457
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1502..1573
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1799..2387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1879..1898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2887..4096
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3933..3952
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4100..5128
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5155..5203
                     /note="synthetic polyadenylation signal"
     CDS             join(5469..5537,5538..6329)
                     /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(5687..5706)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6342..6478
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6623..6694
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6699..6745)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6763..6811
                     /note="synthetic polyadenylation signal"
     enhancer        6892..7195
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7197..7399
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7350..7370
                     /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 cgcaccatca ccatcaccat taaggatccg caggcctctg
      961 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1021 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1081 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1141 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1201 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1261 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     1321 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1381 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1441 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1501 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     1561 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     1621 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     1681 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     1741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2401 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2461 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2521 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2581 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     2641 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     2701 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     2761 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     2821 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     2881 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     2941 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3001 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3061 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3121 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3181 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     3241 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     3301 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     3361 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     3421 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     3481 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     3541 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     3601 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     3661 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     3721 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     3781 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     3841 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     3901 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     3961 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4021 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4081 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     4141 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     4201 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     4261 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     4321 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     4381 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4441 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     4501 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4561 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4621 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     4681 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4741 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4801 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     4861 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     4921 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4981 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     5041 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     5101 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     5161 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     5221 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     5281 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     5341 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     5401 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     5461 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5521 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5581 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5641 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5701 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5761 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5821 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5881 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5941 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6001 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     6061 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     6121 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6181 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6241 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6301 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     6361 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     6421 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     6481 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     6541 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     6601 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     6661 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     6721 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     6781 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     6841 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6901 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6961 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7021 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7081 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7141 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     7201 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7261 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7321 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7381 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     7441 atccactcga cacacccgcc agcggccgc
//
