LOCUS       sequence-295154-        7523 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 7523)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7523)
  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-m4133273
FEATURES             Location/Qualifiers
     source          1..7523
                     /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..994
                     /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"
     polyA_signal    1063..1184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1074..1093)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1128..1147
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1299..1345
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1375..1511
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1556..1627
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1853..2441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1933..1952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2941..4150
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3987..4006
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4154..5182
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5209..5257
                     /note="synthetic polyadenylation signal"
     CDS             join(5523..5591,5592..6383)
                     /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(5741..5760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6396..6532
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6677..6748
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6753..6799)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6817..6865
                     /note="synthetic polyadenylation signal"
     enhancer        6946..7249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7251..7453
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7404..7424
                     /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 cagtcgaaga ggcgatggaa aaagaatttt atcgcggtga gtgcagccaa
      961 cagattcaaa aagatatcct cgtctggtgc cctctaagga tccgcaggcc tctgctagct
     1021 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
     1081 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1141 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1201 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1261 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
     1321 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
     1381 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     1441 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     1501 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     1561 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     1621 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     1681 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     1741 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     1801 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2041 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2281 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2521 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2641 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     2701 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     2761 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     2821 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     2881 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     2941 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     3001 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     3061 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     3121 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     3181 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     3241 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     3301 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     3361 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     3421 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     3481 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     3541 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     3601 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     3661 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     3721 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     3781 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     3841 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     3901 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     3961 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     4021 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     4081 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     4141 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     4201 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     4261 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     4321 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     4381 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     4441 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     4501 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     4561 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     4621 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     4681 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     4741 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     4801 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4861 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4921 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     4981 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     5041 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     5101 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     5161 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     5221 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     5281 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     5341 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     5401 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     5461 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     5521 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5581 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5641 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5701 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5761 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5821 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5881 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5941 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6001 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6061 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     6121 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     6181 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6241 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6301 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6361 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     6421 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     6481 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt
     6541 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     6601 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     6661 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     6721 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     6781 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     6841 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     6901 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6961 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7021 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7081 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     7141 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     7201 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     7261 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     7321 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     7381 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     7441 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     7501 tcgacacacc cgccagcggc cgc
//
