LOCUS       sequence-312900-        8951 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 8951)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8951)
  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-m4134473
FEATURES             Location/Qualifiers
     source          1..8951
                     /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"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3376..3396
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3397..3438
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3491..3612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3521)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3556..3575
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3727..3773
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3803..3939
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3984..4055
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4281..4869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4361..4380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5369..6578
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6415..6434
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6582..7181
                     /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(6582..6601)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7078..7098
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7208..7256
                     /note="synthetic polyadenylation signal"
     CDS             join(7522..7590,7591..8382)
                     /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(7740..7759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8395..8531
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8683..8754
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8759..8805)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8823..8871
                     /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 ccatggctga tgcacaatcg
      601 tttttaaacc gggtttgcgg tgtaagtgca gcccgtctta caccgtgcgg cacaggcact
      661 agtactgatg tcgtatacag ggcttttgac atctacaatg ataaagtagc tggttttgct
      721 aaattcctaa aaactaattg ttgtcgcttc caagaaaagg acgaagatga caatttaatt
      781 gattcttact ttgtagttaa gagacacact ttctctaact accaacatga agaaacaatt
      841 tataatttac ttaaggattg tccagctgtt gctaaacatg acttctttaa gtttagaata
      901 gacggtgaca tggtaccaca tatatcacgt caacgtctta ctaaatacac aatggcagac
      961 ctcgtctatg ctttaaggca ttttgatgaa ggtaattgtg acacattaaa agaaatactt
     1021 gtcacataca attgttgtga tgatgattat ttcaataaaa aggactggta tgattttgta
     1081 gaaaacccag atatattacg cgtatacgcc aacttaggtg aacgtgtacg ccaagctttg
     1141 ttaaaaacag tacaattctg tgatgccatg cgaaatgctg gtattgttgg tgtactgaca
     1201 ttagataatc aagatctcaa tggtaactgg tatgatttcg gtgatttcat acaaaccacg
     1261 ccaggtagtg gagttcctgt tgtagattct tattattcat tgttaatgcc tatattaacc
     1321 ttgaccaggg ctttaactgc agagtcacat gttgacactg acttaacaaa gccttacatt
     1381 aagtgggatt tgttaaaata tgacttcacg gaagagaggt taaaactctt tgaccgttat
     1441 tttaaatatt gggatcagac ataccaccca aattgtgtta actgtttgga tgacagatgc
     1501 attctgcatt gtgcaaactt taatgtttta ttctctacag tgttcccacc tacaagtttt
     1561 ggaccactag tgagaaaaat atttgttgat ggtgttccat ttgtagtttc aactggatac
     1621 cacttcagag agctaggtgt tgtacataat caggatgtaa acttacatag ctctagactt
     1681 agttttaagg aattacttgt gtatgctgct gaccctgcta tgcacgctgc ttctggtaat
     1741 ctattactag ataaacgcac tacgtgcttt tcagtagctg cacttactaa caatgttgct
     1801 tttcaaactg tcaaacccgg taattttaac aaagacttct atgactttgc tgtgtctaag
     1861 ggtttcttta aggaaggaag ttctgttgaa ttaaaacact tcttctttgc tcaggatggt
     1921 aatgctgcta tcagcgatta tgactactat cgttataatc taccaacaat gtgtgatatc
     1981 agacaactac tatttgtagt tgaagttgtt gataagtact ttgattgtta cgatggtggc
     2041 tgtattaatg ctaaccaagt catcgtcaac aacctagaca aatcagctgg ttttccattt
     2101 aataaatggg gtaaggctag actttattat gattcaatga gttatgagga tcaagatgca
     2161 cttttcgcat atacaaaacg taatgtcatc cctactataa ctcaaatgaa tcttaagtat
     2221 gccattagtg caaagaatag agctcgcacc gtagctggtg tctctatctg tagtactatg
     2281 accaatagac agtttcatca aaaattattg aaatcaatag ccgccactag aggagctact
     2341 gtagtaattg gaacaagcaa attctatggt ggttggcaca acatgttaaa aactgtttat
     2401 agtgatgtag aaaaccctca ccttatgggt tgggattatc ctaaatgtga tagagccatg
     2461 cctaacatgc ttagaattat ggcctcactt gttcttgctc gcaaacatac aacgtgttgt
     2521 agcttgtcac accgtttcta tagattagct aatgagtgtg ctcaagtatt gagtgaaatg
     2581 gtcatgtgtg gcggttcact atatgttaaa ccaggtggaa cctcatcagg agatgccaca
     2641 actgcttatg ctaatagtgt ttttaacatt tgtcaagctg tcacggccaa tgttaatgca
     2701 cttttatcta ctgatggtaa caaaattgcc gataagtatg tccgcaattt acaacacaga
     2761 ctttatgagt gtctctatag aaatagagat gttgacacag actttgtgaa tgagttttac
     2821 gcatatttgc gtaaacattt ctcaatgatg atactctctg acgatgctgt tgtgtgtttc
     2881 aatagcactt atgcatctca aggtctagtg gctagcataa agaactttaa gtcagttctt
     2941 tattatcaaa acaatgtttt tatgtctgaa gcaaaatgtt ggactgagac tgaccttact
     3001 aaaggacctc atgaattttg ctctcaacat acaatgctag ttaaacaggg tgatgattat
     3061 gtgtaccttc cttacccaga tccatcaaga atcctagggg ccggctgttt tgtagatgat
     3121 atcgtaaaaa cagatggtac acttatgatt gaacggttcg tgtctttagc tatagatgct
     3181 tacccactta ctaaacatcc taatcaggag tatgctgatg tctttcattt gtacttacaa
     3241 tacataagaa agctacatga tgagttaaca ggacacatgt tagacatgta ttctgttatg
     3301 cttactaatg ataacacttc aaggtattgg gaacctgagt tttatgaggc tatgtacaca
     3361 ccgcatacag tcttagaaaa cctttacttt cagtcgggga aacctatacc caatccactt
     3421 ctcggcttag attctaccta agctagcttg actgactgag atacagcgta ccttcagctc
     3481 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3541 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3601 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3661 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
     3721 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
     3781 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     3841 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     3901 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
     3961 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     4021 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     4081 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
     4141 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
     4201 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4681 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5101 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     5161 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     5221 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     5281 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     5341 cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg
     5401 ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg
     5461 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     5521 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     5581 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     5641 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     5701 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     5761 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     5821 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     5881 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     5941 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     6001 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga
     6061 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     6121 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     6181 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     6241 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     6301 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     6361 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     6421 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     6481 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt
     6541 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca
     6601 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     6661 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     6721 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     6781 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     6841 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     6901 aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     6961 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     7021 gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     7081 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     7141 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt
     7201 cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     7261 acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat
     7321 atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc
     7381 agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga
     7441 tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca
     7501 ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt
     7561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     7621 gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag
     7681 atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg
     7741 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     7801 cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat
     7861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7921 aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     8041 gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact
     8101 ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa
     8161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8221 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     8281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8341 cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg
     8401 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     8461 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     8521 tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     8581 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     8641 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     8701 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
     8761 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
     8821 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg
     8881 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
     8941 atggagttcc g
//
