LOCUS       sequence-295485-        7392 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 7392)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7392)
  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-m4133763
FEATURES             Location/Qualifiers
     source          1..7392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2798..2818
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3167..3184
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3261..3382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3272..3291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3326..3345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3497..3543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3573..3709
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3754..3825
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4051..4639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4131..4150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5385..5453,5454..6245)
                     /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(5603..5622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6258..6394
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6546..6617
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6622..6668)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6686..6734
                     /note="synthetic polyadenylation signal"
     enhancer        6815..7118
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7120..7322
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7273..7293
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7391..7392,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa ccgggtttgc ggtgtaagtg cagcccgtct
       61 tacaccgtgc ggcacaggca ctagtactga tgtcgtatac agggcttttg acatctacaa
      121 tgataaagta gctggttttg ctaaattcct aaaaactaat tgttgtcgct tccaagaaaa
      181 ggacgaagat gacaatttaa ttgattctta ctttgtagtt aagagacaca ctttctctaa
      241 ctaccaacat gaagaaacaa tttataattt acttaaggat tgtccagctg ttgctaaaca
      301 tgacttcttt aagtttagaa tagacggtga catggtacca catatatcac gtcaacgtct
      361 tactaaatac acaatggcag acctcgtcta tgctttaagg cattttgatg aaggtaattg
      421 tgacacatta aaagaaatac ttgtcacata caattgttgt gatgatgatt atttcaataa
      481 aaaggactgg tatgattttg tagaaaaccc agatatatta cgcgtatacg ccaacttagg
      541 tgaacgtgta cgccaagctt tgttaaaaac agtacaattc tgtgatgcca tgcgaaatgc
      601 tggtattgtt ggtgtactga cattagataa tcaagatctc aatggtaact ggtatgattt
      661 cggtgatttc atacaaacca cgccaggtag tggagttcct gttgtagatt cttattattc
      721 attgttaatg cctatattaa ccttgaccag ggctttaact gcagagtcac atgttgacac
      781 tgacttaaca aagccttaca ttaagtggga tttgttaaaa tatgacttca cggaagagag
      841 gttaaaactc tttgaccgtt attttaaata ttgggatcag acataccacc caaattgtgt
      901 taactgtttg gatgacagat gcattctgca ttgtgcaaac tttaatgttt tattctctac
      961 agtgttccca cctacaagtt ttggaccact agtgagaaaa atatttgttg atggtgttcc
     1021 atttgtagtt tcaactggat accacttcag agagctaggt gttgtacata atcaggatgt
     1081 aaacttacat agctctagac ttagttttaa ggaattactt gtgtatgctg ctgaccctgc
     1141 tatgcacgct gcttctggta atctattact agataaacgc actacgtgct tttcagtagc
     1201 tgcacttact aacaatgttg cttttcaaac tgtcaaaccc ggtaatttta acaaagactt
     1261 ctatgacttt gctgtgtcta agggtttctt taaggaagga agttctgttg aattaaaaca
     1321 cttcttcttt gctcaggatg gtaatgctgc tatcagcgat tatgactact atcgttataa
     1381 tctaccaaca atgtgtgata tcagacaact actatttgta gttgaagttg ttgataagta
     1441 ctttgattgt tacgatggtg gctgtattaa tgctaaccaa gtcatcgtca acaacctaga
     1501 caaatcagct ggttttccat ttaataaatg gggtaaggct agactttatt atgattcaat
     1561 gagttatgag gatcaagatg cacttttcgc atatacaaaa cgtaatgtca tccctactat
     1621 aactcaaatg aatcttaagt atgccattag tgcaaagaat agagctcgca ccgtagctgg
     1681 tgtctctatc tgtagtacta tgaccaatag acagtttcat caaaaattat tgaaatcaat
     1741 agccgccact agaggagcta ctgtagtaat tggaacaagc aaattctatg gtggttggca
     1801 caacatgtta aaaactgttt atagtgatgt agaaaaccct caccttatgg gttgggatta
     1861 tcctaaatgt gatagagcca tgcctaacat gcttagaatt atggcctcac ttgttcttgc
     1921 tcgcaaacat acaacgtgtt gtagcttgtc acaccgtttc tatagattag ctaatgagtg
     1981 tgctcaagta ttgagtgaaa tggtcatgtg tggcggttca ctatatgtta aaccaggtgg
     2041 aacctcatca ggagatgcca caactgctta tgctaatagt gtttttaaca tttgtcaagc
     2101 tgtcacggcc aatgttaatg cacttttatc tactgatggt aacaaaattg ccgataagta
     2161 tgtccgcaat ttacaacaca gactttatga gtgtctctat agaaatagag atgttgacac
     2221 agactttgtg aatgagtttt acgcatattt gcgtaaacat ttctcaatga tgatactctc
     2281 tgacgatgct gttgtgtgtt tcaatagcac ttatgcatct caaggtctag tggctagcat
     2341 aaagaacttt aagtcagttc tttattatca aaacaatgtt tttatgtctg aagcaaaatg
     2401 ttggactgag actgacctta ctaaaggacc tcatgaattt tgctctcaac atacaatgct
     2461 agttaaacag ggtgatgatt atgtgtacct tccttaccca gatccatcaa gaatcctagg
     2521 ggccggctgt tttgtagatg atatcgtaaa aacagatggt acacttatga ttgaacggtt
     2581 cgtgtcttta gctatagatg cttacccact tactaaacat cctaatcagg agtatgctga
     2641 tgtctttcat ttgtacttac aatacataag aaagctacat gatgagttaa caggacacat
     2701 gttagacatg tattctgtta tgcttactaa tgataacact tcaaggtatt gggaacctga
     2761 gttttatgag gctatgtaca caccgcatac agtcttagaa aacctttact ttcagtcgat
     2821 gagtgacaaa atcattcatc taaccgatga ctcctttgat acagacgtgt tgaaggccga
     2881 tggtgcgata ctggtagatt tctgggctga atggtgtggc ccgtgcaaga tgatcgcacc
     2941 cattctggac gagatcgccg acgagtatca aggaaaatta actgtcgcta agcttaatat
     3001 agatcagaac ccagggacgg cacctaaata cggcatcagg ggaataccta ccctcctgct
     3061 ctttaagaat ggggaagttg ccgcgacaaa agtgggagcc ctgtctaagg gccagctaaa
     3121 agagttcttg gacgctaacc tggcaggttc aggctcgggg cacatgcacc atcaccatca
     3181 ccatagcagc gggctcgtgc cacgcggcta agctagcttg actgactgag atacagcgta
     3241 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3421 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     3481 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     3541 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     3601 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     3661 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     3721 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     3781 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     3841 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     3901 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     3961 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     4021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4861 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     4921 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     4981 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     5041 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     5101 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     5161 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5221 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5281 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5341 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     5401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     5521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     5581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5761 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5941 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     6001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6241 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     6301 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     6361 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     6421 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     6481 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     6541 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     6601 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     6661 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     6721 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     6781 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6841 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6901 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6961 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     7021 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     7081 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     7141 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     7201 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     7261 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     7321 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     7381 gccagcggcc gc
//
