LOCUS       sequence-318500-        5127 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 5127)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5127)
  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-m4134315
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1474..1494
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1495..1521
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1574..1695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1585..1604)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1639..1658
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1810..1856
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1886..2022
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2067..2138
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2364..2952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2444..2463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3698..3766,3767..4558)
                     /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(3916..3935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4571..4707
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4859..4930
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4935..4981)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4999..5047
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagtca agcgtggcaa
      601 ccgggtgttg ctatgcctaa tctttacaaa atgcaaagaa tgctattaga aaagtgtgac
      661 cttcaaaatt atggtgatag tgcaacatta cctaaaggca taatgatgaa tgtcgcaaaa
      721 tatactcaac tgtgtcaata tttaaacaca ttaacattag ctgtacccta taatatgaga
      781 gttatacatt ttggtgctgg ttctgataaa ggagttgcac caggtacagc tgttttaaga
      841 cagtggttgc ctacgggtac gctgcttgtc gattcagatc ttaatgactt tgtctctgat
      901 gcagattcaa ctttgattgg tgattgtgca actgtacata cagctaataa atgggatctc
      961 attattagtg atatgtacga ccctaagact aaaaatgtta caaaagaaaa tgactctaaa
     1021 gagggttttt tcacttacat ttgtgggttt atacaacaaa agctagctct tggaggttcc
     1081 gtggctataa agataacaga acattcttgg aatgctgatc tttataagct catgggacac
     1141 ttcgcatggt ggacagcctt tgttactaat gtgaatgcgt catcatctga agcattttta
     1201 attggatgta attatcttgg caaaccacgc gaacaaatag atggttatgt catgcatgca
     1261 aattacatat tttggaggaa tacaaatcca attcagttgt cttcctattc tttatttgac
     1321 atgagtaaat ttccccttaa attaaggggt actgctgtta tgtctttaaa agaaggtcaa
     1381 atcaatgata tgattttatc tcttcttagt aaaggtagac ttataattag agaaaacaac
     1441 agagttgtta tttctagtga tgttcttgtt aacgaaaacc tttactttca gtcgtacccc
     1501 tatgacgtac cagattatgc gtaagctagc ttgactgact gagatacagc gtaccttcag
     1561 ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     1621 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1681 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1741 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta
     1801 tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc
     1861 cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct
     1921 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     1981 ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc
     2041 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
     2101 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
     2161 atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     2221 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     2281 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2341 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2401 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2461 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2521 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2581 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2641 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2701 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2761 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2821 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2881 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2941 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3001 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3061 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3121 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3181 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     3241 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     3301 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     3361 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     3421 atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg
     3481 agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc
     3541 aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt
     3601 taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa
     3661 catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt
     3721 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3781 ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga
     3841 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag
     3901 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3961 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga
     4021 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4081 tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc
     4141 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4201 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt
     4261 gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg
     4321 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4381 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4441 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4501 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc
     4561 gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     4621 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     4681 tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct
     4741 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     4801 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     4861 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     4921 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     4981 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     5041 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     5101 agttcatagc ccatatatgg agttccg
//
