LOCUS       sequence-320321-        7082 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 7082)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7082)
  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-m4133288
FEATURES             Location/Qualifiers
     source          1..7082
                     /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             1480..1500
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1504..1569
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1622..1743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1633..1652)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1687..1706
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1858..1904
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1934..2070
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2115..2186
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2412..3000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2492..2511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3500..4709
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4546..4565
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4713..5312
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4713..4732)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5209..5229
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5339..5387
                     /note="synthetic polyadenylation signal"
     CDS             join(5653..5721,5722..6513)
                     /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(5871..5890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6526..6662
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6814..6885
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6890..6936)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6954..7002
                     /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 ccatgtcctc acaagcgtgg
      601 cagccaggcg tagcaatgcc taatttatac aagatgcagc ggatgctcct tgaaaaatgc
      661 gacctacaga actatggaga tagtgctaca ttgcccaagg ggataatgat gaatgtggcc
      721 aaatataccc aactgtgtca gtacctgaac actctcacgc tggccgttcc gtacaacatg
      781 agggtgattc attttggtgc cggcagcgac aagggagtcg ctcccgggac cgctgtgctg
      841 cgacagtggc ttcctacagg cactctgcta gtggattctg atttgaatga cttcgtcagc
      901 gacgcagact cgaccctcat cggagattgt gcgactgttc acacagccaa caaatgggac
      961 ctgatcattt ccgatatgta tgatccaaag accaaaaatg tcacgaagga gaacgactca
     1021 aaggagggtt tctttacgta catctgcggg ttcatccaac agaaacttgc actgggcgga
     1081 agtgtggcaa ttaaaataac agaacactct tggaacgccg acctatacaa gcttatgggt
     1141 cattttgctt ggtggaccgc ctttgttact aatgtaaacg cttcaagctc cgaggccttc
     1201 ctgataggtt gtaattattt aggcaagcct agagagcaga ttgatgggta tgtgatgcac
     1261 gcaaattaca tcttctggcg caacacaaat ccaattcagc tcagttctta ttcactgttt
     1321 gatatgagca aattccccct taaactgaga ggcactgccg tcatgtcctt gaaggaagga
     1381 caaatcaacg atatgatctt gtctctcctt agcaaggggc gtttgattat tagggagaac
     1441 aatcgggtgg tgatctcatc ggacgttctg gtcaacaatg aaaaccttta ctttcagtcg
     1501 atggattata aggaccacga tggagactac aaagatcatg acattgatta caaggacgac
     1561 gatgacaaat aagctagctt gactgactga gatacagcgt accttcagct cacagacatg
     1621 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1681 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1741 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1801 ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag
     1861 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga
     1921 ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     1981 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     2041 gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc
     2101 gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta
     2161 gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa
     2221 ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag
     2281 aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt
     2341 ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
     3241 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
     3301 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
     3361 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
     3421 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
     3481 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
     3541 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
     3601 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
     3661 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
     3721 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
     3781 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
     3841 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
     3901 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
     3961 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     4021 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     4081 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     4141 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     4201 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     4261 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     4321 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     4381 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     4441 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     4501 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     4561 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     4621 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     4681 tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc acggtgcgcc
     4741 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4801 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     4861 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     4921 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4981 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5041 agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5101 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5161 ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5221 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5281 cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag tcttagataa
     5341 taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc
     5401 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     5461 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     5521 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     5581 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     5641 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5701 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5761 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5821 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
     5881 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5941 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     6001 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6061 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6121 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6181 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
     6241 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
     6301 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6361 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6421 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6481 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     6541 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     6601 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     6661 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     6721 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     6781 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     6841 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     6901 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     6961 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     7021 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     7081 cg
//
