LOCUS       sequence-300401-        6981 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 6981)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6981)
  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-m4135860
FEATURES             Location/Qualifiers
     source          1..6981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      125..261
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    413..484
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(489..535)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    553..601
                     /note="synthetic polyadenylation signal"
     enhancer        682..985
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        987..1189
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1140..1160
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1258..1267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4109..4230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4120..4139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4174..4193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4345..4391
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4421..4557
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4602..4673
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4899..5487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4979..4998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6233..6301,6302..6981,1..112)
                     /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(6451..6470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
       61 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
      121 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
      181 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
      241 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
      301 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
      361 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
      421 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
      481 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
      541 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
      601 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
      661 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      721 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      781 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      841 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      901 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      961 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     1021 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1081 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1141 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     1201 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgcc
     1261 accatggctg atgcacaatc gtttttaaac cgggtttgcg gtgtaagtgc agcccgtctt
     1321 acaccgtgcg gcacaggcac tagtactgat gtcgtataca gggcttttga catctacaat
     1381 gataaagtag ctggttttgc taaattccta aaaactaatt gttgtcgctt ccaagaaaag
     1441 gacgaagatg acaatttaat tgattcttac tttgtagtta agagacacac tttctctaac
     1501 taccaacatg aagaaacaat ttataattta cttaaggatt gtccagctgt tgctaaacat
     1561 gacttcttta agtttagaat agacggtgac atggtaccac atatatcacg tcaacgtctt
     1621 actaaataca caatggcaga cctcgtctat gctttaaggc attttgatga aggtaattgt
     1681 gacacattaa aagaaatact tgtcacatac aattgttgtg atgatgatta tttcaataaa
     1741 aaggactggt atgattttgt agaaaaccca gatatattac gcgtatacgc caacttaggt
     1801 gaacgtgtac gccaagcttt gttaaaaaca gtacaattct gtgatgccat gcgaaatgct
     1861 ggtattgttg gtgtactgac attagataat caagatctca atggtaactg gtatgatttc
     1921 ggtgatttca tacaaaccac gccaggtagt ggagttcctg ttgtagattc ttattattca
     1981 ttgttaatgc ctatattaac cttgaccagg gctttaactg cagagtcaca tgttgacact
     2041 gacttaacaa agccttacat taagtgggat ttgttaaaat atgacttcac ggaagagagg
     2101 ttaaaactct ttgaccgtta ttttaaatat tgggatcaga cataccaccc aaattgtgtt
     2161 aactgtttgg atgacagatg cattctgcat tgtgcaaact ttaatgtttt attctctaca
     2221 gtgttcccac ctacaagttt tggaccacta gtgagaaaaa tatttgttga tggtgttcca
     2281 tttgtagttt caactggata ccacttcaga gagctaggtg ttgtacataa tcaggatgta
     2341 aacttacata gctctagact tagttttaag gaattacttg tgtatgctgc tgaccctgct
     2401 atgcacgctg cttctggtaa tctattacta gataaacgca ctacgtgctt ttcagtagct
     2461 gcacttacta acaatgttgc ttttcaaact gtcaaacccg gtaattttaa caaagacttc
     2521 tatgactttg ctgtgtctaa gggtttcttt aaggaaggaa gttctgttga attaaaacac
     2581 ttcttctttg ctcaggatgg taatgctgct atcagcgatt atgactacta tcgttataat
     2641 ctaccaacaa tgtgtgatat cagacaacta ctatttgtag ttgaagttgt tgataagtac
     2701 tttgattgtt acgatggtgg ctgtattaat gctaaccaag tcatcgtcaa caacctagac
     2761 aaatcagctg gttttccatt taataaatgg ggtaaggcta gactttatta tgattcaatg
     2821 agttatgagg atcaagatgc acttttcgca tatacaaaac gtaatgtcat ccctactata
     2881 actcaaatga atcttaagta tgccattagt gcaaagaata gagctcgcac cgtagctggt
     2941 gtctctatct gtagtactat gaccaataga cagtttcatc aaaaattatt gaaatcaata
     3001 gccgccacta gaggagctac tgtagtaatt ggaacaagca aattctatgg tggttggcac
     3061 aacatgttaa aaactgttta tagtgatgta gaaaaccctc accttatggg ttgggattat
     3121 cctaaatgtg atagagccat gcctaacatg cttagaatta tggcctcact tgttcttgct
     3181 cgcaaacata caacgtgttg tagcttgtca caccgtttct atagattagc taatgagtgt
     3241 gctcaagtat tgagtgaaat ggtcatgtgt ggcggttcac tatatgttaa accaggtgga
     3301 acctcatcag gagatgccac aactgcttat gctaatagtg tttttaacat ttgtcaagct
     3361 gtcacggcca atgttaatgc acttttatct actgatggta acaaaattgc cgataagtat
     3421 gtccgcaatt tacaacacag actttatgag tgtctctata gaaatagaga tgttgacaca
     3481 gactttgtga atgagtttta cgcatatttg cgtaaacatt tctcaatgat gatactctct
     3541 gacgatgctg ttgtgtgttt caatagcact tatgcatctc aaggtctagt ggctagcata
     3601 aagaacttta agtcagttct ttattatcaa aacaatgttt ttatgtctga agcaaaatgt
     3661 tggactgaga ctgaccttac taaaggacct catgaatttt gctctcaaca tacaatgcta
     3721 gttaaacagg gtgatgatta tgtgtacctt ccttacccag atccatcaag aatcctaggg
     3781 gccggctgtt ttgtagatga tatcgtaaaa acagatggta cacttatgat tgaacggttc
     3841 gtgtctttag ctatagatgc ttacccactt actaaacatc ctaatcagga gtatgctgat
     3901 gtctttcatt tgtacttaca atacataaga aagctacatg atgagttaac aggacacatg
     3961 ttagacatgt attctgttat gcttactaat gataacactt caaggtattg ggaacctgag
     4021 ttttatgagg ctatgtacac accgcataca gtcttataag ctagcttgac tgactgagat
     4081 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     4141 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4201 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4261 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4321 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     4381 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     4441 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     4501 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     4561 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     4621 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     4681 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     4741 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     4801 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     4861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5701 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     5761 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     5821 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     5881 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     5941 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg taccgagaac
     6001 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     6061 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     6121 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     6181 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     6241 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6301 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     6361 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6421 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6481 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6541 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6601 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     6661 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6721 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6781 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     6841 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6901 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6961 cattgcagca ctggggccag a
//
