LOCUS       sequence-295269-        7490 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 7490)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7490)
  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-m4134174
FEATURES             Location/Qualifiers
     source          1..7490
                     /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             917..961
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1030..1151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1041..1060)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1095..1114
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1266..1312
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1342..1478
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1523..1594
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1820..2408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1900..1919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2908..4117
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3954..3973
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4121..5149
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5176..5224
                     /note="synthetic polyadenylation signal"
     CDS             join(5490..5558,5559..6350)
                     /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(5708..5727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6363..6499
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6644..6715
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6720..6766)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6784..6832
                     /note="synthetic polyadenylation signal"
     enhancer        6913..7216
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7218..7420
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7371..7391
                     /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 cagtcgaaag aaaccgcggc agctaagttc gagaggcagc acatggacag
      961 ttaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     1021 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1081 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1141 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1201 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     1261 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     1321 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     1381 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     1441 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     1501 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     1561 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     1621 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     1681 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     1741 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1801 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1981 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2221 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2641 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     2701 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     2761 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     2821 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     2881 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     2941 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     3001 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     3061 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     3121 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     3181 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     3241 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     3301 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     3361 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     3421 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     3481 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     3541 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     3601 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     3661 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     3721 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     3781 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     3841 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     3901 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     3961 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     4021 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     4081 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     4141 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4201 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4261 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     4321 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     4381 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     4441 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     4501 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     4561 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     4621 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     4681 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     4741 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     4801 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     4861 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     4921 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     4981 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5041 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     5101 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     5161 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     5221 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     5281 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     5341 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     5401 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     5461 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     5521 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5581 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     5641 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     5701 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5761 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     5821 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5881 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     5941 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6001 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6061 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6121 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6181 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6241 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6301 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     6361 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     6421 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     6481 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     6541 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     6601 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     6661 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     6721 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     6781 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     6841 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     6901 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     6961 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7021 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7081 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7141 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7201 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7261 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     7321 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     7381 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     7441 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
