LOCUS       sequence-295494-        5118 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 5118)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5118)
  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-m4133768
FEATURES             Location/Qualifiers
     source          1..5118
                     /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             917..934
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    987..1108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(998..1017)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1052..1071
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1223..1269
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1299..1435
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1480..1551
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1777..2365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1857..1876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3111..3179,3180..3971)
                     /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(3329..3348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3984..4120
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4272..4343
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4348..4394)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4412..4460
                     /note="synthetic polyadenylation signal"
     enhancer        4541..4844
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4846..5048
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4999..5019
                     /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 cagtcgcacc atcaccatca ccattaagct agcttgactg actgagatac
      961 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
     1021 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     1081 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     1141 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     1201 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     1261 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     1321 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     1381 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     1441 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     1501 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     1561 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     1621 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     1681 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     1741 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2281 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2341 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2401 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2461 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2521 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2581 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     2641 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     2701 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     2761 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     2821 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     2881 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     2941 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     3001 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     3061 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     3121 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3181 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     3241 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     3301 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3361 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     3421 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3481 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     3541 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3601 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3661 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3721 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3781 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3841 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3901 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3961 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     4021 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     4081 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     4141 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     4201 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     4261 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     4321 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     4381 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     4441 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     4501 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4561 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4621 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4681 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4741 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4801 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     4861 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4921 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4981 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     5041 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     5101 acacccgcca gcggccgc
//
