LOCUS       sequence-308753-        5655 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 5655)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5655)
  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-m4133339
FEATURES             Location/Qualifiers
     source          1..5655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1474..1494
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1528..1605
                     /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             1633..1653
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2102..2223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2113..2132)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2167..2186
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2338..2384
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2414..2550
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2595..2666
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2892..3480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2972..2991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4226..4294,4295..5086)
                     /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(4444..4463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5099..5235
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5387..5458
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5463..5509)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5527..5575
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagtca agcgtggcaa
      601 ccgggtgttg ctatgcctaa tctttacaaa atgcaaagaa tgctattaga aaagtgtgac
      661 cttcaaaatt atggtgatag tgcaacatta cctaaaggca taatgatgaa tgtcgcaaaa
      721 tatactcaac tgtgtcaata tttaaacaca ttaacattag ctgtacccta taatatgaga
      781 gttatacatt ttggtgctgg ttctgataaa ggagttgcac caggtacagc tgttttaaga
      841 cagtggttgc ctacgggtac gctgcttgtc gattcagatc ttaatgactt tgtctctgat
      901 gcagattcaa ctttgattgg tgattgtgca actgtacata cagctaataa atgggatctc
      961 attattagtg atatgtacga ccctaagact aaaaatgtta caaaagaaaa tgactctaaa
     1021 gagggttttt tcacttacat ttgtgggttt atacaacaaa agctagctct tggaggttcc
     1081 gtggctataa agataacaga acattcttgg aatgctgatc tttataagct catgggacac
     1141 ttcgcatggt ggacagcctt tgttactaat gtgaatgcgt catcatctga agcattttta
     1201 attggatgta attatcttgg caaaccacgc gaacaaatag atggttatgt catgcatgca
     1261 aattacatat tttggaggaa tacaaatcca attcagttgt cttcctattc tttatttgac
     1321 atgagtaaat ttccccttaa attaaggggt actgctgtta tgtctttaaa agaaggtcaa
     1381 atcaatgata tgattttatc tcttcttagt aaaggtagac ttataattag agaaaacaac
     1441 agagttgtta tttctagtga tgttcttgtt aacgaaaacc tttactttca gtcgggacgg
     1501 cgaatcccgg ggctgataaa tccctggaaa aggagatgga agaagaactt tattgcagtg
     1561 tctgcggcta accggttcaa aaagatcagt tcaagcggcg ccctagatta cgacattcct
     1621 acgaccgcct ccgagaatct ctatttccag ggtgaatttg gcttggcaca acatgacgag
     1681 gccgtcgata acaagtttaa taaagaacag cagaacgctt tctatgagat ccttcactta
     1741 ccaaatctga acgaagagca acgcaacgct ttcattcaga gcctgaaaga cgatccctcg
     1801 cagtctgcaa atctcctggc cgaagccaag aagctgaatg acgcacaggc tccaaaagtt
     1861 gataacaagt ttaacaagga gcagcagaat gctttctacg agatattgca ccttcctaat
     1921 ctgaatgagg aacaaagaaa tgcatttatc cagtccctaa aagacgatcc atcacagagt
     1981 gccaacctgc ttgcggaagc caaaaagctc aatgatgccc aggccccgaa agtagacgcg
     2041 aaccatcaat aagctagctt gactgactga gatacagcgt accttcagct cacagacatg
     2101 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2161 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2221 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2281 ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag
     2341 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga
     2401 ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     2461 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     2521 gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc
     2581 gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta
     2641 gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa
     2701 ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag
     2761 aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt
     2821 ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3601 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3661 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
     3721 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
     3781 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
     3841 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
     3901 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
     3961 tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4021 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     4081 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4141 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     4201 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     4261 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4321 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     4381 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     4441 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4501 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     4561 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4621 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     4681 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4741 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     4801 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     4861 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4921 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4981 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5041 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     5101 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5161 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     5221 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     5281 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     5341 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     5401 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     5461 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     5521 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     5581 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     5641 atatatggag ttccg
//
