LOCUS       sequence-295384-        7390 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 7390)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7390)
  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-m4133173
FEATURES             Location/Qualifiers
     source          1..7390
                     /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"
     promoter        3237..4446
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4283..4302
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4450..5049
                     /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(4450..4469)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4946..4966
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5076..5124
                     /note="synthetic polyadenylation signal"
     CDS             join(5390..5458,5459..6250)
                     /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(5608..5627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6263..6399
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6544..6615
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6620..6666)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6684..6732
                     /note="synthetic polyadenylation signal"
     enhancer        6813..7116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7118..7320
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7271..7291
                     /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 gcgcctggcc
     3241 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     3301 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     3361 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     3421 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     3481 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     3541 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     3601 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     3661 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     3721 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     3781 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     3841 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     3901 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     3961 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     4021 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     4081 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     4141 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     4201 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     4261 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     4321 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     4381 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     4441 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4501 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4561 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4621 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4681 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4741 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     4801 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4861 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     4921 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4981 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5041 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     5101 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     5161 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     5221 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     5281 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     5341 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     5401 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5461 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     5521 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     5581 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5641 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     5701 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5761 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     5821 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5881 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5941 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6001 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6061 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6121 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6181 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6241 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     6301 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     6361 cttccatacg tgtcctcctt accagaaatt tatccttaag atcgtttaaa ctcgactctg
     6421 gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg
     6481 tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc
     6541 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
     6601 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     6661 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
     6721 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
     6781 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     6841 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     6901 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6961 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     7021 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     7081 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
     7141 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     7201 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     7261 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     7321 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
     7381 cagcggccgc
//
