LOCUS       sequence-295779-        7841 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 7841)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7841)
  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-m4133335
FEATURES             Location/Qualifiers
     source          1..7841
                     /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             1271..1288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1381..1502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1392..1411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1446..1465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1617..1663
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1693..1829
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1874..1945
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2171..2759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2251..2270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3259..4468
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4305..4324
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4472..5500
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5527..5575
                     /note="synthetic polyadenylation signal"
     CDS             join(5841..5909,5910..6701)
                     /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(6059..6078)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6714..6850
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6995..7066
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7071..7117)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7135..7183
                     /note="synthetic polyadenylation signal"
     enhancer        7264..7567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7569..7771
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7722..7742
                     /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 cgatgagtga caaaatcatt catctaaccg atgactcctt
      961 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
     1021 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
     1081 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
     1141 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
     1201 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
     1261 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaaggatc
     1321 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     1381 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1501 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1561 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1621 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     1681 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1741 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1801 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     1861 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1921 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     1981 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     2041 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     2101 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2161 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2221 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2281 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2341 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2401 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2461 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2521 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2581 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2641 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2701 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2761 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2821 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2881 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2941 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     3001 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     3061 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     3121 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     3181 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     3241 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     3301 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     3361 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3421 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3481 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3541 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3601 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     3661 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     3721 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     3781 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     3841 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3901 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     3961 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     4021 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     4081 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     4141 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     4201 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     4261 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     4321 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     4381 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     4441 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     4501 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4561 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4621 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4681 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     4741 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4801 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     4861 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4921 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4981 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     5041 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     5101 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     5161 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     5221 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     5281 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     5341 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     5401 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     5461 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     5521 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     5581 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     5641 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     5701 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     5761 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     5821 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     5881 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5941 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6001 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     6061 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6121 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     6181 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6241 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6301 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6361 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     6421 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     6481 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6541 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6601 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6661 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     6721 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     6781 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     6841 ttatccttaa gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     6901 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     6961 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     7021 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     7081 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     7141 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     7201 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7261 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     7321 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     7381 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     7441 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     7501 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     7561 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     7621 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     7681 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     7741 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     7801 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
