LOCUS       sequence-295751-        7021 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 7021)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7021)
  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-m4133237
FEATURES             Location/Qualifiers
     source          1..7021
                     /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"
     promoter        2868..4077
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3914..3933
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4081..4680
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4081..4100)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4577..4597
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4707..4755
                     /note="synthetic polyadenylation signal"
     CDS             join(5021..5089,5090..5881)
                     /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(5239..5258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5894..6030
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6175..6246
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6251..6297)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6315..6363
                     /note="synthetic polyadenylation signal"
     enhancer        6444..6747
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6749..6951
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6902..6922
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7020..7021,1..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 cgcgcctggc ctccgcgccg
     2881 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     2941 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     3001 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     3061 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     3121 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     3181 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     3241 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     3301 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     3361 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     3421 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     3481 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     3541 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     3601 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     3661 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     3721 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     3781 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     3841 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     3901 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     3961 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     4021 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     4081 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     4141 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     4201 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4261 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4321 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4381 tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag
     4441 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4501 agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg
     4561 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4621 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag
     4681 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     4741 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     4801 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     4861 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     4921 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     4981 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     5041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     5161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     5221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     5341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5401 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     5461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5581 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     5641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5881 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     5941 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     6001 gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct ggctctatcg
     6061 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     6121 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     6181 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     6241 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     6301 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     6361 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     6421 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6481 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6541 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6601 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6661 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     6721 tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt
     6781 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6841 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6901 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     6961 aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg
     7021 c
//
