LOCUS       sequence-317271-        7029 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 7029)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7029)
  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-m4134155
FEATURES             Location/Qualifiers
     source          1..7029
                     /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..3423
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3476..3597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3487..3506)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3541..3560
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3712..3758
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3788..3924
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3969..4040
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4266..4854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4346..4365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5600..5668,5669..6460)
                     /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(5818..5837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6473..6609
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6761..6832
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6837..6883)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6901..6949
                     /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 cagtcgtacc cctatgacgt accagattat
     3421 gcgtaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     3481 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3541 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3601 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3661 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     3721 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     3781 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     3841 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     3901 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     3961 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     4021 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     4081 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     4141 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     4201 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4921 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4981 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5041 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     5101 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     5161 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     5221 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     5281 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     5341 tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     5401 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     5461 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     5521 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     5581 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     5641 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5701 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5761 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     5821 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5881 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5941 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6001 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6061 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6121 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6181 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     6241 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6301 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6361 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6421 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     6481 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     6541 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     6601 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6661 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6721 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     6781 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6841 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6901 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     6961 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     7021 ggagttccg
//
