LOCUS       sequence-295950-        7398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7398)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7398)
  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-m4134361
FEATURES             Location/Qualifiers
     source          1..7398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2804..2824
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3173..3190
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3267..3388
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3278..3297)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3332..3351
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3503..3549
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3579..3715
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3760..3831
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4057..4645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4137..4156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5391..5459,5460..6251)
                     /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(5609..5628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6264..6400
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6552..6623
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6628..6674)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6692..6740
                     /note="synthetic polyadenylation signal"
     enhancer        6821..7124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7126..7328
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7279..7299
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct gccgacgcac agtcgttcct gaatagagtt tgtggtgtat ccgcggcccg
       61 gctaactcca tgcggaaccg ggacgagcac agatgtggtg taccgagctt ttgatatcta
      121 taacgacaaa gtcgctggct tcgccaagtt tcttaagacc aactgttgca ggttccaaga
      181 gaaagacgaa gatgataatc tcatagactc atactttgtg gttaagcgtc acacttttag
      241 taattatcag catgaggaaa caatttataa cttgctgaag gactgccctg ccgtagcaaa
      301 acacgacttc tttaaattcc gcatcgatgg cgatatggtc ccgcatatta gcagacagcg
      361 gttaaccaag tacactatgg cagacctggt gtacgccctg cgacacttcg acgagggaaa
      421 ctgtgacacg ttaaaagaaa tcctcgtgac ttataattgt tgcgatgacg attatttcaa
      481 caagaaagat tggtacgatt ttgtcgagaa tcccgatata ctcagagtgt acgctaacct
      541 gggggaaagg gttcgccagg cactattgaa gacagtccaa ttttgcgacg cgatgcggaa
      601 tgccggcatt gtgggtgtcc tgacacttga taaccaggac ctgaatggca actggtatga
      661 ctttggagat ttcattcaga ccacgcccgg gtccggagta cctgtcgttg actcttacta
      721 ctcacttctg atgccaatcc taaccttaac ccgagctctc acagctgaga gtcacgtgga
      781 caccgacctc acaaaaccgt atatcaagtg ggatttgctg aagtacgatt tcacggaaga
      841 gaggttaaag ctctttgatc gttatttcaa atactgggac cagacttatc atcccaactg
      901 tgtgaattgt ctggacgaca gatgcatact tcattgtgcc aatttcaatg tactgtttag
      961 cacagtgttc ccaccaactt cgtttggtcc tctagtgcgc aaaatctttg tggatggggt
     1021 gcccttcgtt gtttccacag gctatcactt tcgtgagctc ggcgtagttc acaaccaaga
     1081 tgtcaacctg cattcttcca ggttaagttt taaggaactt ctggtgtatg ccgcagaccc
     1141 tgcgatgcac gctgcaagcg gtaatttgtt gctggataag cggaccacgt gcttcagcgt
     1201 ggcggcttta acaaataacg ttgccttcca aaccgtcaag cctggcaact tcaataaaga
     1261 tttctacgat ttcgccgtca gtaaagggtt ttttaaggag ggatcaagcg tagaattgaa
     1321 acattttttc tttgctcagg acggtaatgc cgctatctcg gactacgact actaccggta
     1381 taacctacca actatgtgcg acattcggca gctgttgttc gtggtggagg tcgttgataa
     1441 atactttgat tgctacgatg gcggatgcat taacgccaat caagtcatcg tgaacaatct
     1501 ggacaagagc gccggcttcc cctttaacaa atggggaaag gcaaggctat attatgactc
     1561 tatgtcctat gaggatcagg acgctctctt cgcgtatacc aagcgcaacg ttatccccac
     1621 aattacccag atgaacctca aatatgcaat cagcgcaaag aacagagcga ggacagttgc
     1681 tggggtgagt atatgttcga caatgactaa caggcagttt caccagaagc tgttgaaatc
     1741 tattgccgct acacgagggg ctaccgtcgt gattggcacc tccaaattct acggagggtg
     1801 gcataatatg ttaaaaactg tgtactccga cgtggaaaac cctcacctga tgggttggga
     1861 ttatcctaag tgcgaccggg cgatgccaaa catgctgaga attatggcgt cattagtcct
     1921 ggcgcgaaag cacaccacgt gctgtagtct gtcacatcgc ttttacagat tggctaatga
     1981 atgcgcccaa gtgcttagcg agatggtgat gtgtgggggt tcactttacg tgaagcccgg
     2041 aggaacgtcc tcaggcgacg ccacaactgc ctacgcaaat tcagtattca acatatgtca
     2101 ggccgttaca gcaaacgtga atgctctgct gtctactgat gggaacaaga tcgccgataa
     2161 gtacgtccgg aacctgcaac acaggctgta cgagtgtctg tatcgtaatc gagatgtaga
     2221 caccgacttc gtgaacgaat tttacgcata cttacggaaa catttctcca tgatgatcct
     2281 ctccgacgat gcagttgtgt gctttaattc aacttacgcc tctcagggct tggtggccag
     2341 tatcaaaaat tttaagtcgg ttctttatta tcagaataac gtgtttatga gcgaggctaa
     2401 gtgttggacc gagactgatc tgaccaaagg cccgcacgag ttctgttccc agcacaccat
     2461 gctggtcaag cagggtgatg actatgtcta tttaccatac ccagatccga gtaggatctt
     2521 gggagccggt tgttttgtgg atgacattgt gaagactgac ggtacactca tgattgagcg
     2581 cttcgtcagt ttggcaatcg acgcttaccc ccttaccaaa catccaaatc aggaatatgc
     2641 ggatgtgttt cacctgtacc ttcaatacat aaggaaactt catgatgaac taactggaca
     2701 tatgctcgac atgtattctg taatgcttac aaatgacaat acctcacggt actgggaacc
     2761 ggagttttat gaagccatgt acaccccaca tactgtcctg caagaaaacc tttactttca
     2821 gtcgatgagt gacaaaatca ttcatctaac cgatgactcc tttgatacag acgtgttgaa
     2881 ggccgatggt gcgatactgg tagatttctg ggctgaatgg tgtggcccgt gcaagatgat
     2941 cgcacccatt ctggacgaga tcgccgacga gtatcaagga aaattaactg tcgctaagct
     3001 taatatagat cagaacccag ggacggcacc taaatacggc atcaggggaa tacctaccct
     3061 cctgctcttt aagaatgggg aagttgccgc gacaaaagtg ggagccctgt ctaagggcca
     3121 gctaaaagag ttcttggacg ctaacctggc aggttcaggc tcggggcaca tgcaccatca
     3181 ccatcaccat agcagcgggc tcgtgccacg cggctaagct agcttgactg actgagatac
     3241 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
     3301 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3361 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3421 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3481 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     3541 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     3601 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     3661 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     3721 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     3781 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     3841 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     3901 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     3961 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     4021 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4081 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4141 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4201 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4261 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4321 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4381 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4441 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4501 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4561 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4621 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4741 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4801 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4861 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     4921 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     4981 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     5041 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     5101 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     5161 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     5221 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     5281 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     5341 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     5401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     5521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     5581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     5701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     5821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5941 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6001 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6241 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     6301 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     6361 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     6421 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     6481 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     6541 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     6601 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     6661 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     6721 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     6781 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6841 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6901 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6961 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     7021 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     7081 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     7141 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     7201 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     7261 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     7321 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     7381 acacccgcca gcggccgc
//
