LOCUS       sequence-295479-        5175 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 5175)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5175)
  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-m4133839
FEATURES             Location/Qualifiers
     source          1..5175
                     /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             926..991
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1044..1165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1055..1074)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1109..1128
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1280..1326
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1356..1492
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1537..1608
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1834..2422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1914..1933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3168..3236,3237..4028)
                     /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(3386..3405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4041..4177
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4329..4400
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4405..4451)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4469..4517
                     /note="synthetic polyadenylation signal"
     enhancer        4598..4901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4903..5105
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5056..5076
                     /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 cgatggatta taaggaccac gatggagact acaaagatca
      961 tgacattgat tacaaggacg acgatgacaa ataagctagc ttgactgact gagatacagc
     1021 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1081 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1141 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1201 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1261 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1321 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1381 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1441 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1501 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1561 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1621 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1681 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1741 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2341 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2401 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2461 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2521 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2581 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2641 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2701 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2761 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2821 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2881 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     2941 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     3001 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     3061 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     3121 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     3181 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3241 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     3301 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     3361 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3421 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     3481 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3541 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     3601 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3661 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3721 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     3781 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3841 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3901 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3961 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4021 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     4081 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     4141 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     4201 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     4261 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     4321 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     4381 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     4441 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     4501 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     4561 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4621 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4681 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4741 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4801 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4861 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     4921 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4981 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     5041 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     5101 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     5161 cccgccagcg gccgc
//
