LOCUS       sequence-444320-        8007 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 8007)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8007)
  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-m4133279
FEATURES             Location/Qualifiers
     source          1..8007
                     /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             950..1027
                     /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             1055..1075
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1540..1661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1551..1570)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1605..1624
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1776..1822
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1852..1988
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2033..2104
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2330..2918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2410..2429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3418..4627
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4464..4483
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4631..5659
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5686..5734
                     /note="synthetic polyadenylation signal"
     CDS             join(6000..6068,6069..6860)
                     /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(6218..6237)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6873..7009
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7161..7232
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7237..7283)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7301..7349
                     /note="synthetic polyadenylation signal"
     enhancer        7430..7733
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7735..7937
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7888..7908
                     /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 cagtcgggac ggcgaatccc ggggctgata aatccctgga aaaggagatg
      961 gaagaagaac tttattgcag tgtctgcggc taaccggttc aaaaagatca gttcaagcgg
     1021 cgccctagat tacgacattc ctacgaccgc ctccgagaat ctctatttcc agggtgaatt
     1081 tggcttggca caacatgacg aggccgtcga taacaagttt aataaagaac agcagaacgc
     1141 tttctatgag atccttcact taccaaatct gaacgaagag caacgcaacg ctttcattca
     1201 gagcctgaaa gacgatccct cgcagtctgc aaatctcctg gccgaagcca agaagctgaa
     1261 tgacgcacag gctccaaaag ttgataacaa gtttaacaag gagcagcaga atgctttcta
     1321 cgagatattg caccttccta atctgaatga ggaacaaaga aatgcattta tccagtccct
     1381 aaaagacgat ccatcacaga gtgccaacct gcttgcggaa gccaaaaagc tcaatgatgc
     1441 ccaggccccg aaagtagacg cgaaccatca ataaggatcc gcaggcctct gctagcttga
     1501 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
     1561 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1621 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1681 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1741 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
     1801 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
     1861 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     1921 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     1981 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     2041 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     2101 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     2161 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     2221 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     2281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2521 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2581 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2641 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2701 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2761 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2821 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2881 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2941 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3001 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3061 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3121 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     3181 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     3241 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     3301 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     3361 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc
     3421 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca
     3481 cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     3541 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     3601 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     3661 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     3721 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     3781 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     3841 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     3901 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     3961 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     4021 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct
     4081 aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt
     4141 gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg
     4201 cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac
     4261 gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta
     4321 ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca
     4381 ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt
     4441 atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc
     4501 gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg
     4561 taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt
     4621 gttagacaac atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     4681 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4741 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4801 tttctacaaa gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga
     4861 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4921 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4981 ggaggcaatg gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt
     5041 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     5101 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     5161 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     5221 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     5281 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     5341 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     5401 tgcaggatcg ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     5461 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     5521 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt
     5581 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     5641 tccgagggca aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc
     5701 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     5761 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     5821 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     5881 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     5941 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     6001 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6061 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     6121 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6181 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6241 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6301 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6361 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     6421 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6481 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6541 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     6601 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6661 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6721 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6781 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6841 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     6901 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     6961 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     7021 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     7081 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     7141 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     7201 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     7261 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     7321 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     7381 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     7441 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     7501 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     7561 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     7621 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     7681 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     7741 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     7801 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     7861 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     7921 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     7981 ccactcgaca cacccgccag cggccgc
//
