LOCUS       sequence-295843-        5490 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 5490)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5490)
  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-m4133208
FEATURES             Location/Qualifiers
     source          1..5490
                     /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             1265..1282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1359..1480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1370..1389)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1424..1443
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1595..1641
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1671..1807
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1852..1923
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2149..2737)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2229..2248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3483..3551,3552..4343)
                     /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(3701..3720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4356..4492
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4644..4715
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4720..4766)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4784..4832
                     /note="synthetic polyadenylation signal"
     enhancer        4913..5216
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5218..5420
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5371..5391
                     /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 cagtcgatga gtgacaaaat cattcatcta accgatgact cctttgatac
      961 agacgtgttg aaggccgatg gtgcgatact ggtagatttc tgggctgaat ggtgtggccc
     1021 gtgcaagatg atcgcaccca ttctggacga gatcgccgac gagtatcaag gaaaattaac
     1081 tgtcgctaag cttaatatag atcagaaccc agggacggca cctaaatacg gcatcagggg
     1141 aatacctacc ctcctgctct ttaagaatgg ggaagttgcc gcgacaaaag tgggagccct
     1201 gtctaagggc cagctaaaag agttcttgga cgctaacctg gcaggttcag gctcggggca
     1261 catgcaccat caccatcacc atagcagcgg gctcgtgcca cgcggctaag ctagcttgac
     1321 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     1381 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1441 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1501 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1561 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     1621 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     1681 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     1741 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     1801 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     1861 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1921 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1981 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     2041 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     2101 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2161 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2221 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2281 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2341 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2401 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2461 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2521 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2581 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2641 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2701 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2761 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2821 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2881 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2941 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     3001 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     3061 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     3121 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     3181 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     3241 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     3301 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     3361 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     3421 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     3481 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3541 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3601 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3661 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3721 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3781 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3841 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3901 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3961 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4021 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     4081 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     4141 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4201 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4261 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4321 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     4381 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     4441 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     4501 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     4561 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     4621 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     4681 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     4741 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     4801 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     4861 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4921 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4981 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5041 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5101 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5161 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     5221 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     5281 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     5341 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     5401 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     5461 catccactcg acacacccgc cagcggccgc
//
