LOCUS       sequence-295077-        9431 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 9431)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9431)
  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-m4134303
FEATURES             Location/Qualifiers
     source          1..9431
                     /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             2825..2902
                     /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"
     polyA_signal    2971..3092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2982..3001)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3036..3055
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3207..3253
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3283..3419
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3464..3535
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3761..4349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3841..3860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4849..6058
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5895..5914
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6062..7090
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    7117..7165
                     /note="synthetic polyadenylation signal"
     CDS             join(7431..7499,7500..8291)
                     /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(7649..7668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8304..8440
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8585..8656
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8661..8707)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8725..8773
                     /note="synthetic polyadenylation signal"
     enhancer        8854..9157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9159..9361
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9312..9332
                     /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 gtcgaagagg cgatggaaaa agaattttat cgcggtgagt gcagccaaca gattcaaaaa
     2881 gatatcctcg tctggtgccc tctaaggatc cgcaggcctc tgctagcttg actgactgag
     2941 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
     3001 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3061 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3121 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3181 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
     3241 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
     3301 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     3361 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     3421 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
     3481 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
     3541 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     3601 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     3661 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     3721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4201 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4261 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4321 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4381 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4441 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4501 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4561 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     4621 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     4681 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     4741 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     4801 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
     4861 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
     4921 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     4981 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     5041 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     5101 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     5161 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     5221 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     5281 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     5341 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     5401 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     5461 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
     5521 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
     5581 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     5641 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     5701 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
     5761 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     5821 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     5881 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     5941 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     6001 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
     6061 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     6121 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     6181 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     6241 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga
     6301 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     6361 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat
     6421 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     6481 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     6541 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     6601 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     6661 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     6721 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     6781 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     6841 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     6901 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     6961 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga
     7021 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     7081 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga
     7141 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga
     7201 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     7261 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     7321 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     7381 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     7441 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7501 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     7561 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     7621 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7681 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     7741 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7801 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     7861 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7921 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7981 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8041 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8101 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     8161 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8221 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8281 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     8341 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     8401 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct
     8461 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     8521 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     8581 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     8641 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     8701 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     8761 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     8821 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     8881 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     8941 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     9001 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     9061 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     9121 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     9181 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     9241 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     9301 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     9361 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     9421 ccagcggccg c
//
