LOCUS       sequence-295259-        5121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5121)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5121)
  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-m4134101
FEATURES             Location/Qualifiers
     source          1..5121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             872..892
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             893..937
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    990..1111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1001..1020)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1055..1074
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1226..1272
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1302..1438
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1483..1554
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1780..2368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1860..1879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3114..3182,3183..3974)
                     /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(3332..3351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3987..4123
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4275..4346
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4351..4397)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4415..4463
                     /note="synthetic polyadenylation signal"
     enhancer        4544..4847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4849..5051
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5002..5022
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5120..5121,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gtgaaaagaa caatcaaggg tacacaccac tggttgttac tcacaatttt
       61 gacttcactt ttagttttag tccagagtac tcaatggtct ttgttctttt ttttgtatga
      121 aaatgccttt ttaccttttg ctatgggtat tattgctatg tctgcttttg caatgatgtt
      181 tgtcaaacat aagcatgcat ttctctgttt gtttttgtta ccttctcttg ccactgtagc
      241 ttattttaat atggtctata tgcctgctag ttgggtgatg cgtattatga catggttgga
      301 tatggttgat actagtttgt ctggttttaa gctaaaagac tgtgttatgt atgcatcagc
      361 tgtagtgtta ctaatcctta tgacagcaag aactgtgtat gatgatggtg ctaggagagt
      421 gtggacactt atgaatgtct tgacactcgt ttataaagtt tattatggta atgctttaga
      481 tcaagccatt tccatgtggg ctcttataat ctctgttact tctaactact caggtgtagt
      541 tacaactgtc atgtttttgg ccagaggtat tgtttttatg tgtgttgagt attgccctat
      601 tttcttcata actggtaata cacttcagtg tataatgcta gtttattgtt tcttaggcta
      661 tttttgtact tgttactttg gcctcttttg tttactcaac cgctacttta gactgactct
      721 tggtgtttat gattacttag tttctacaca ggagtttaga tatatgaatt cacagggact
      781 actcccaccc aagaatagca tagatgcctt caaactcaac attaaattgt tgggtgttgg
      841 tggcaaacct tgtatcaaag tagccactgt agaaaacctt tactttcagt cgaaagaaac
      901 cgcggcagct aagttcgaga ggcagcacat ggacagttaa gctagcttga ctgactgaga
      961 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1021 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1081 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1141 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1201 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1261 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1321 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1381 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1441 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1501 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1561 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1621 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1681 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1741 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1801 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1861 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1921 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1981 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2041 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2101 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2161 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2221 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2281 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2401 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2461 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2521 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2581 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2641 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2701 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2761 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2821 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2881 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2941 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     3001 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3061 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     3121 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3181 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3241 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3301 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3361 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3421 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3481 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3541 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3601 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3661 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3721 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3781 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3841 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3901 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3961 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     4021 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4081 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     4141 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4201 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     4261 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     4321 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     4381 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     4441 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     4501 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4561 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4621 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4681 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4741 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4801 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4861 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4921 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4981 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     5041 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     5101 gacacacccg ccagcggccg c
//
