LOCUS       sequence-487616-        7077 bp    DNA     circular SYN 01-OCT-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 7077)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7077)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134347
FEATURES             Location/Qualifiers
     source          1..7077
                     /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             2828..2893
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2946..3067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2957..2976)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3011..3030
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3182..3228
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3258..3394
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3439..3510
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3736..4324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3816..3835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5070..5138,5139..5930)
                     /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(5288..5307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5943..6079
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6231..6302
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6307..6353)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6371..6419
                     /note="synthetic polyadenylation signal"
     enhancer        6500..6803
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6805..7007
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6958..6978
                     /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 gtcgatggat tataaggacc acgatggaga ctacaaagat catgacattg attacaagga
     2881 cgacgatgac aaataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     2941 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3001 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3061 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3121 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     3181 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     3241 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     3301 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     3361 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     3421 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     3481 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     3541 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     3601 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     3661 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4561 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     4621 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     4681 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     4741 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     4801 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     4861 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     4921 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     4981 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     5041 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     5101 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5161 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     5221 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     5281 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5341 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     5401 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5461 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     5521 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5581 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     5641 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     5701 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5761 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5821 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5881 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     5941 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     6001 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     6061 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     6121 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     6181 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     6241 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     6301 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     6361 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     6421 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     6481 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6541 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     6601 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6661 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6721 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6781 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     6841 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6901 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     6961 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     7021 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
