LOCUS       sequence-295684-        7027 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 7027)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7027)
  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-m4133229
FEATURES             Location/Qualifiers
     source          1..7027
                     /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..943
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    996..1117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1007..1026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1061..1080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1232..1278
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1308..1444
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1489..1560
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1786..2374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1866..1885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2874..4083
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3920..3939
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4087..4686
                     /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(4087..4106)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4583..4603
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4713..4761
                     /note="synthetic polyadenylation signal"
     CDS             join(5027..5095,5096..5887)
                     /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(5245..5264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5900..6036
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6181..6252
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6257..6303)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6321..6369
                     /note="synthetic polyadenylation signal"
     enhancer        6450..6753
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6755..6957
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6908..6928
                     /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 cagtcgtacc cctatgacgt accagattat gcgtaagcta gcttgactga
      961 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     1021 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1081 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1141 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1201 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     1261 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     1321 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     1381 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     1441 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     1501 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     1561 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     1621 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     1681 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2581 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     2641 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     2701 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2761 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2821 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     2881 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     2941 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     3001 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     3061 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     3121 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     3181 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     3241 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     3301 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     3361 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     3421 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     3481 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     3541 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     3601 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     3661 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     3721 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     3781 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     3841 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     3901 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     3961 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     4021 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     4081 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4141 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4201 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4261 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4321 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4381 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     4441 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4501 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     4561 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4621 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4681 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     4741 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     4801 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     4861 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     4921 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     4981 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     5041 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5101 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     5161 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     5221 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5281 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     5341 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5401 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     5461 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5521 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5581 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     5641 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5701 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5761 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5821 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5881 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     5941 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     6001 ccatacgtgt cctccttacc agaaatttat ccttaagatc gtttaaactc gactctggct
     6061 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     6121 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     6181 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     6241 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     6301 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     6361 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     6421 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     6481 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     6541 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6601 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6661 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6721 tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat
     6781 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     6841 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     6901 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     6961 ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag
     7021 cggccgc
//
