LOCUS       sequence-369826-        9470 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 9470)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9470)
  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-m4134323
FEATURES             Location/Qualifiers
     source          1..9470
                     /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             2858..2935
                     /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             2963..2983
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3432..3553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3497..3516
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3668..3714
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3744..3880
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3925..3996
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4222..4810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4302..4321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5310..6519
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6356..6375
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6523..7122
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6523..6542)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7019..7039
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7149..7197
                     /note="synthetic polyadenylation signal"
     CDS             join(7463..7531,7532..8323)
                     /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(7681..7700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8336..8472
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8624..8695
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8700..8746)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8764..8812
                     /note="synthetic polyadenylation signal"
     enhancer        8893..9196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9198..9400
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9351..9371
                     /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 gtcgggacgg cgaatcccgg ggctgataaa tccctggaaa aggagatgga agaagaactt
     2881 tattgcagtg tctgcggcta accggttcaa aaagatcagt tcaagcggcg ccctagatta
     2941 cgacattcct acgaccgcct ccgagaatct ctatttccag ggtgaatttg gcttggcaca
     3001 acatgacgag gccgtcgata acaagtttaa taaagaacag cagaacgctt tctatgagat
     3061 ccttcactta ccaaatctga acgaagagca acgcaacgct ttcattcaga gcctgaaaga
     3121 cgatccctcg cagtctgcaa atctcctggc cgaagccaag aagctgaatg acgcacaggc
     3181 tccaaaagtt gataacaagt ttaacaagga gcagcagaat gctttctacg agatattgca
     3241 ccttcctaat ctgaatgagg aacaaagaaa tgcatttatc cagtccctaa aagacgatcc
     3301 atcacagagt gccaacctgc ttgcggaagc caaaaagctc aatgatgccc aggccccgaa
     3361 agtagacgcg aaccatcaat aagctagctt gactgactga gatacagcgt accttcagct
     3421 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3481 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3541 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3601 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     3661 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     3721 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     3781 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     3841 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     3901 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     3961 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     4021 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     4081 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     4141 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4201 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4261 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4321 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4381 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4441 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4501 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4561 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4621 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4681 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4741 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4921 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4981 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5041 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     5101 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     5161 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     5221 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     5281 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     5341 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     5401 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     5461 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     5521 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     5581 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     5641 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     5701 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     5761 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     5821 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     5881 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     5941 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     6001 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     6061 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     6121 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     6181 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     6241 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     6301 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     6361 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     6421 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     6481 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     6541 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     6601 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     6661 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     6721 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     6781 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     6841 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     6901 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     6961 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     7021 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     7081 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     7141 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     7201 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     7261 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     7321 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     7381 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     7441 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7501 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7561 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     7621 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     7681 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7741 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     7801 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7861 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     7921 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7981 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     8041 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     8101 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8161 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     8221 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8281 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     8341 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     8401 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     8461 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     8521 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     8581 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     8641 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     8701 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     8761 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     8821 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     8881 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     8941 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     9001 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     9061 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     9121 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     9181 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     9241 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     9301 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     9361 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     9421 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
