LOCUS       sequence-297885-        7010 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 7010)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7010)
  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-m4134157
FEATURES             Location/Qualifiers
     source          1..7010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      273..282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1146..1166
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1167..1193
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1246..1367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1257..1276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1311..1330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1482..1528
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1558..1694
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1739..1810
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2036..2624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2116..2135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3124..4333
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4170..4189
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4337..4936
                     /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(4337..4356)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4833..4853
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4963..5011
                     /note="synthetic polyadenylation signal"
     CDS             join(5277..5345,5346..6137)
                     /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(5495..5514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6150..6286
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6438..6509
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6514..6560)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6578..6626
                     /note="synthetic polyadenylation signal"
     enhancer        6707..7010
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
      241 ctaccatcca ctcgacacac ccgccagcgg ccgccaccat ggcagtgaaa agaacaatca
      301 agggtacaca ccactggttg ttactcacaa ttttgacttc acttttagtt ttagtccaga
      361 gtactcaatg gtctttgttc ttttttttgt atgaaaatgc ctttttacct tttgctatgg
      421 gtattattgc tatgtctgct tttgcaatga tgtttgtcaa acataagcat gcatttctct
      481 gtttgttttt gttaccttct cttgccactg tagcttattt taatatggtc tatatgcctg
      541 ctagttgggt gatgcgtatt atgacatggt tggatatggt tgatactagt ttgtctggtt
      601 ttaagctaaa agactgtgtt atgtatgcat cagctgtagt gttactaatc cttatgacag
      661 caagaactgt gtatgatgat ggtgctagga gagtgtggac acttatgaat gtcttgacac
      721 tcgtttataa agtttattat ggtaatgctt tagatcaagc catttccatg tgggctctta
      781 taatctctgt tacttctaac tactcaggtg tagttacaac tgtcatgttt ttggccagag
      841 gtattgtttt tatgtgtgtt gagtattgcc ctattttctt cataactggt aatacacttc
      901 agtgtataat gctagtttat tgtttcttag gctatttttg tacttgttac tttggcctct
      961 tttgtttact caaccgctac tttagactga ctcttggtgt ttatgattac ttagtttcta
     1021 cacaggagtt tagatatatg aattcacagg gactactccc acccaagaat agcatagatg
     1081 ccttcaaact caacattaaa ttgttgggtg ttggtggcaa accttgtatc aaagtagcca
     1141 ctgtagaaaa cctttacttt cagtcgtacc cctatgacgt accagattat gcgtaagcta
     1201 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
     1261 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1321 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1381 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1441 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
     1501 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
     1561 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
     1621 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
     1681 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
     1741 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     1801 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
     1861 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     1921 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     1981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2461 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     2881 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     2941 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     3001 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     3061 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     3121 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     3181 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     3241 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     3301 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     3361 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     3421 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     3481 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     3541 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     3601 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     3661 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     3721 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     3781 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     3841 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     3901 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     3961 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt
     4021 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     4081 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     4141 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     4201 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     4261 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     4321 ttttttgtta gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4381 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4441 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4501 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4561 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4621 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc
     4681 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4741 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg
     4801 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4861 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4921 caagcccggt gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat
     4981 cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga
     5041 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     5101 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     5161 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     5221 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     5281 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5341 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     5401 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5461 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5521 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5581 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5641 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     5701 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5761 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5821 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     5881 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5941 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6001 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6061 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6121 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     6181 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     6241 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     6301 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     6361 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     6421 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     6481 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     6541 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     6601 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     6661 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     6721 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     6781 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     6841 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     6901 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     6961 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
