LOCUS       sequence-297935-        5133 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 5133)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5133)
  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-m4133218
FEATURES             Location/Qualifiers
     source          1..5133
                     /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..949
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1002..1123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1013..1032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1067..1086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1238..1284
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1314..1450
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1495..1566
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1792..2380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1872..1891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3126..3194,3195..3986)
                     /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(3344..3363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3999..4135
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4287..4358
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4363..4409)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4427..4475
                     /note="synthetic polyadenylation signal"
     enhancer        4556..4859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4861..5063
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5014..5034
                     /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 cgtaccccta tgacgtacca gattatgcgt aagctagctt
      961 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
     1021 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1081 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1141 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1201 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     1261 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     1321 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     1381 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     1441 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     1501 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1561 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1621 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1681 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2221 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2461 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2521 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2581 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2641 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2701 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2761 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2821 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2881 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     2941 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     3001 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     3061 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     3121 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3241 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3301 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3421 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3541 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3661 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     3721 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     3781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3961 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     4021 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     4081 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     4141 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     4201 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     4261 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     4321 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     4381 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     4441 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     4501 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4561 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4621 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4681 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4741 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4801 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4861 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4921 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4981 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5041 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     5101 taccatccac tcgacacacc cgccagcggc cgc
//
