LOCUS       sequence-308756-        7026 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 7026)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7026)
  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-m4134288
FEATURES             Location/Qualifiers
     source          1..7026
                     /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"
     CDS             3382..3402
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3403..3420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3473..3594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3484..3503)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3538..3557
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3709..3755
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3785..3921
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3966..4037
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4263..4851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4343..4362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5597..5665,5666..6457)
                     /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(5815..5834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6470..6606
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6758..6829
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6834..6880)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6898..6946
                     /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 ccatgtctgc cgacgcacag
      601 tcgttcctga atagagtttg tggtgtatcc gcggcccggc taactccatg cggaaccggg
      661 acgagcacag atgtggtgta ccgagctttt gatatctata acgacaaagt cgctggcttc
      721 gccaagtttc ttaagaccaa ctgttgcagg ttccaagaga aagacgaaga tgataatctc
      781 atagactcat actttgtggt taagcgtcac acttttagta attatcagca tgaggaaaca
      841 atttataact tgctgaagga ctgccctgcc gtagcaaaac acgacttctt taaattccgc
      901 atcgatggcg atatggtccc gcatattagc agacagcggt taaccaagta cactatggca
      961 gacctggtgt acgccctgcg acacttcgac gagggaaact gtgacacgtt aaaagaaatc
     1021 ctcgtgactt ataattgttg cgatgacgat tatttcaaca agaaagattg gtacgatttt
     1081 gtcgagaatc ccgatatact cagagtgtac gctaacctgg gggaaagggt tcgccaggca
     1141 ctattgaaga cagtccaatt ttgcgacgcg atgcggaatg ccggcattgt gggtgtcctg
     1201 acacttgata accaggacct gaatggcaac tggtatgact ttggagattt cattcagacc
     1261 acgcccgggt ccggagtacc tgtcgttgac tcttactact cacttctgat gccaatccta
     1321 accttaaccc gagctctcac agctgagagt cacgtggaca ccgacctcac aaaaccgtat
     1381 atcaagtggg atttgctgaa gtacgatttc acggaagaga ggttaaagct ctttgatcgt
     1441 tatttcaaat actgggacca gacttatcat cccaactgtg tgaattgtct ggacgacaga
     1501 tgcatacttc attgtgccaa tttcaatgta ctgtttagca cagtgttccc accaacttcg
     1561 tttggtcctc tagtgcgcaa aatctttgtg gatggggtgc ccttcgttgt ttccacaggc
     1621 tatcactttc gtgagctcgg cgtagttcac aaccaagatg tcaacctgca ttcttccagg
     1681 ttaagtttta aggaacttct ggtgtatgcc gcagaccctg cgatgcacgc tgcaagcggt
     1741 aatttgttgc tggataagcg gaccacgtgc ttcagcgtgg cggctttaac aaataacgtt
     1801 gccttccaaa ccgtcaagcc tggcaacttc aataaagatt tctacgattt cgccgtcagt
     1861 aaagggtttt ttaaggaggg atcaagcgta gaattgaaac attttttctt tgctcaggac
     1921 ggtaatgccg ctatctcgga ctacgactac taccggtata acctaccaac tatgtgcgac
     1981 attcggcagc tgttgttcgt ggtggaggtc gttgataaat actttgattg ctacgatggc
     2041 ggatgcatta acgccaatca agtcatcgtg aacaatctgg acaagagcgc cggcttcccc
     2101 tttaacaaat ggggaaaggc aaggctatat tatgactcta tgtcctatga ggatcaggac
     2161 gctctcttcg cgtataccaa gcgcaacgtt atccccacaa ttacccagat gaacctcaaa
     2221 tatgcaatca gcgcaaagaa cagagcgagg acagttgctg gggtgagtat atgttcgaca
     2281 atgactaaca ggcagtttca ccagaagctg ttgaaatcta ttgccgctac acgaggggct
     2341 accgtcgtga ttggcacctc caaattctac ggagggtggc ataatatgtt aaaaactgtg
     2401 tactccgacg tggaaaaccc tcacctgatg ggttgggatt atcctaagtg cgaccgggcg
     2461 atgccaaaca tgctgagaat tatggcgtca ttagtcctgg cgcgaaagca caccacgtgc
     2521 tgtagtctgt cacatcgctt ttacagattg gctaatgaat gcgcccaagt gcttagcgag
     2581 atggtgatgt gtgggggttc actttacgtg aagcccggag gaacgtcctc aggcgacgcc
     2641 acaactgcct acgcaaattc agtattcaac atatgtcagg ccgttacagc aaacgtgaat
     2701 gctctgctgt ctactgatgg gaacaagatc gccgataagt acgtccggaa cctgcaacac
     2761 aggctgtacg agtgtctgta tcgtaatcga gatgtagaca ccgacttcgt gaacgaattt
     2821 tacgcatact tacggaaaca tttctccatg atgatcctct ccgacgatgc agttgtgtgc
     2881 tttaattcaa cttacgcctc tcagggcttg gtggccagta tcaaaaattt taagtcggtt
     2941 ctttattatc agaataacgt gtttatgagc gaggctaagt gttggaccga gactgatctg
     3001 accaaaggcc cgcacgagtt ctgttcccag cacaccatgc tggtcaagca gggtgatgac
     3061 tatgtctatt taccataccc agatccgagt aggatcttgg gagccggttg ttttgtggat
     3121 gacattgtga agactgacgg tacactcatg attgagcgct tcgtcagttt ggcaatcgac
     3181 gcttaccccc ttaccaaaca tccaaatcag gaatatgcgg atgtgtttca cctgtacctt
     3241 caatacataa ggaaacttca tgatgaacta actggacata tgctcgacat gtattctgta
     3301 atgcttacaa atgacaatac ctcacggtac tgggaaccgg agttttatga agccatgtac
     3361 accccacata ctgtcctgca agaaaacctt tactttcagt cgcaccatca ccatcaccat
     3421 taagctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     3481 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3541 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3601 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3661 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     3721 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     3781 accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     3841 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     3901 ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat
     3961 caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     4021 actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt
     4081 caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc
     4141 agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa
     4201 atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4261 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4381 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4441 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4501 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4561 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4621 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4681 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4741 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4801 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4861 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4921 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4981 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5041 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg
     5101 aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc
     5161 tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg
     5221 gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag
     5281 agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg
     5341 gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     5401 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     5461 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     5521 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     5581 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5701 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5761 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     5821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5881 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     5941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6001 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6121 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     6181 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     6241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6421 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     6481 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     6541 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     6601 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     6661 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     6721 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     6781 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     6841 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     6901 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     6961 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7021 gttccg
//
