LOCUS       sequence-295336-        7561 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 7561)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7561)
  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-m4133255
FEATURES             Location/Qualifiers
     source          1..7561
                     /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             956..1033
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1061..1081
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1530..1651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1541..1560)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1595..1614
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1766..1812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1842..1978
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2023..2094
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2320..2908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2400..2419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3408..4617
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4454..4473
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4621..5220
                     /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(4621..4640)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5117..5137
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5247..5295
                     /note="synthetic polyadenylation signal"
     CDS             join(5561..5629,5630..6421)
                     /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(5779..5798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6434..6570
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6715..6786
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6791..6837)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6855..6903
                     /note="synthetic polyadenylation signal"
     enhancer        6984..7287
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7289..7491
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7442..7462
                     /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 cgggacggcg aatcccgggg ctgataaatc cctggaaaag
      961 gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc
     1021 aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct atttccaggg
     1081 tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata aagaacagca
     1141 gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac gcaacgcttt
     1201 cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg aagccaagaa
     1261 gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc agcagaatgc
     1321 tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg catttatcca
     1381 gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca aaaagctcaa
     1441 tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga ctgactgaga
     1501 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1561 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1621 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1681 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1741 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1801 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1861 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1921 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1981 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     2041 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     2101 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     2161 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     2221 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     2281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3121 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     3181 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     3241 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     3301 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3361 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg
     3421 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     3481 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     3541 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     3601 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     3661 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     3721 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     3781 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     3841 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     3901 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     3961 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     4021 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     4081 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     4141 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     4201 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     4261 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     4321 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     4381 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     4441 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     4501 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     4561 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     4621 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     4681 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     4741 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4801 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4861 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4921 tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag
     4981 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5041 agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg
     5101 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5161 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag
     5221 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     5281 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     5341 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     5401 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     5461 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     5521 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     5581 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5641 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     5701 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     5761 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5821 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     5881 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5941 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     6001 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6061 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6121 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     6181 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6241 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6301 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6361 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6421 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     6481 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     6541 gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct ggctctatcg
     6601 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     6661 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     6721 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     6781 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     6841 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     6901 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     6961 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     7021 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     7081 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     7141 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     7201 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     7261 tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt
     7321 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     7381 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     7441 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     7501 aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg
     7561 c
//
