LOCUS       sequence-295132-        9743 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 9743)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9743)
  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-m4134242
FEATURES             Location/Qualifiers
     source          1..9743
                     /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             3173..3190
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3283..3404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3294..3313)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3348..3367
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3519..3565
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3595..3731
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3776..3847
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4073..4661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4153..4172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5161..6370
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6207..6226
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6374..7402
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    7429..7477
                     /note="synthetic polyadenylation signal"
     CDS             join(7743..7811,7812..8603)
                     /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(7961..7980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8616..8752
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8897..8968
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8973..9019)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9037..9085
                     /note="synthetic polyadenylation signal"
     enhancer        9166..9469
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9471..9673
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9624..9644
                     /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 gtcgatgagt gacaaaatca ttcatctaac cgatgactcc tttgatacag acgtgttgaa
     2881 ggccgatggt gcgatactgg tagatttctg ggctgaatgg tgtggcccgt gcaagatgat
     2941 cgcacccatt ctggacgaga tcgccgacga gtatcaagga aaattaactg tcgctaagct
     3001 taatatagat cagaacccag ggacggcacc taaatacggc atcaggggaa tacctaccct
     3061 cctgctcttt aagaatgggg aagttgccgc gacaaaagtg ggagccctgt ctaagggcca
     3121 gctaaaagag ttcttggacg ctaacctggc aggttcaggc tcggggcaca tgcaccatca
     3181 ccatcaccat agcagcgggc tcgtgccacg cggctaagga tccgcaggcc tctgctagct
     3241 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
     3301 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3361 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3421 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3481 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
     3541 taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg
     3601 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     3661 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     3721 tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc
     3781 ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc
     3841 ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct
     3901 acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc
     3961 gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag
     4021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4501 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4861 tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa
     4921 ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga
     4981 acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta
     5041 tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa
     5101 cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct
     5161 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg
     5221 ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc
     5281 ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg acattttagg
     5341 acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga
     5401 aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg
     5461 attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc
     5521 gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg
     5581 gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc
     5641 aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc
     5701 agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag
     5761 gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc
     5821 gctaatgcgg gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga
     5881 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     5941 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     6001 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     6061 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     6121 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     6181 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     6241 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
     6301 atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt
     6361 tttgttagac aacatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     6421 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     6481 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     6541 tggtttctac aaagatcgtt atgtttatcg gcactttgca tccgccgcgc tcccgattcc
     6601 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     6661 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     6721 cgcggaggca atggatgcca tcgctgccgc cgatcttagc cagacgagcg ggttcggccc
     6781 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     6841 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     6901 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     6961 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     7021 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     7081 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     7141 gcttgcagga tcgccccggc tccgggcgta tatgctccgc attggtcttg accaactcta
     7201 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     7261 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgccgc
     7321 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     7381 tcgtccgagg gcaaaggaat aagctagtat gtaagcctag tcttagataa taaaatcgct
     7441 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg
     7501 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     7561 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     7621 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     7681 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     7741 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7801 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7861 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7921 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7981 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     8041 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     8101 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     8161 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     8221 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     8281 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     8341 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     8401 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     8461 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     8521 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     8581 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     8641 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     8701 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt
     8761 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     8821 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     8881 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     8941 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     9001 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     9061 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     9121 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     9181 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     9241 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     9301 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     9361 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     9421 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     9481 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     9541 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     9601 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     9661 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     9721 tcgacacacc cgccagcggc cgc
//
