LOCUS       sequence-295080-        5655 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 5655)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5655)
  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-m4133339
FEATURES             Location/Qualifiers
     source          1..5655
                     /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             950..1027
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1055..1075
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1524..1645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1535..1554)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1589..1608
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1760..1806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1836..1972
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2017..2088
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2314..2902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2394..2413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3648..3716,3717..4508)
                     /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(3866..3885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4521..4657
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4809..4880
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4885..4931)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4949..4997
                     /note="synthetic polyadenylation signal"
     enhancer        5078..5381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5383..5585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5536..5556
                     /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 cagtcgggac ggcgaatccc ggggctgata aatccctgga aaaggagatg
      961 gaagaagaac tttattgcag tgtctgcggc taaccggttc aaaaagatca gttcaagcgg
     1021 cgccctagat tacgacattc ctacgaccgc ctccgagaat ctctatttcc agggtgaatt
     1081 tggcttggca caacatgacg aggccgtcga taacaagttt aataaagaac agcagaacgc
     1141 tttctatgag atccttcact taccaaatct gaacgaagag caacgcaacg ctttcattca
     1201 gagcctgaaa gacgatccct cgcagtctgc aaatctcctg gccgaagcca agaagctgaa
     1261 tgacgcacag gctccaaaag ttgataacaa gtttaacaag gagcagcaga atgctttcta
     1321 cgagatattg caccttccta atctgaatga ggaacaaaga aatgcattta tccagtccct
     1381 aaaagacgat ccatcacaga gtgccaacct gcttgcggaa gccaaaaagc tcaatgatgc
     1441 ccaggccccg aaagtagacg cgaaccatca ataagctagc ttgactgact gagatacagc
     1501 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1561 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1621 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1681 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1741 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1801 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1861 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1921 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1981 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     2041 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     2101 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     2161 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     2221 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     2281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3121 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     3181 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     3241 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3301 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3361 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     3421 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     3481 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     3541 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     3601 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     3661 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3721 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     3781 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     3841 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3901 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     3961 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4021 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     4081 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4141 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4201 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     4261 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4321 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4381 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4441 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4501 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     4561 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     4621 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     4681 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     4741 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     4801 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     4861 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     4921 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     4981 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     5041 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     5101 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     5161 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     5221 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     5281 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     5341 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     5401 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     5461 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     5521 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     5581 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     5641 cccgccagcg gccgc
//
