LOCUS       sequence-317727-        7016 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 7016)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7016)
  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-m4133907
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1456..1476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1477..1503
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1556..1677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1567..1586)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1621..1640
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1792..1838
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1868..2004
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2049..2120
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2346..2934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2426..2445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3434..4643
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4480..4499
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4647..5246
                     /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(4647..4666)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5143..5163
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5273..5321
                     /note="synthetic polyadenylation signal"
     CDS             join(5587..5655,5656..6447)
                     /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(5805..5824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6460..6596
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6748..6819
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6824..6870)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6888..6936
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagcgc cgttaagcgg
      601 acaatcaaag ggacccatca ctggttgctc cttactatac tgacgtcttt actggtcttg
      661 gtacagagta cacaatggtc gcttttcttt tttctgtacg agaatgcatt cctcccattt
      721 gcgatgggaa tcattgctat gtccgccttc gctatgatgt tcgtgaagca caaacatgcc
      781 ttcctttgcc tgtttctcct accctcactg gcaaccgtgg cctattttaa catggtgtac
      841 atgcctgcga gctgggtcat gcgaattatg acttggctgg acatggttga tacctcccta
      901 tctggcttca agttaaagga ctgtgtgatg tatgcatcag ctgtagtgct gctaatcctg
      961 atgacagctc gcacagtcta cgatgacggt gccaggagag tgtggactct gatgaatgtc
     1021 ctcaccttag tttataaagt atactatggc aacgctctcg atcaggccat tagtatgtgg
     1081 gcattgataa tctccgtgac gagtaactac agcggagtgg taaccactgt catgtttttg
     1141 gcacgcggga tagtgttcat gtgtgtcgaa tattgcccga tcttttttat cacagggaat
     1201 actctgcaat gcattatgct ggtttactgt ttcttgggct atttctgcac ctgttatttt
     1261 ggtctattct gcctgcttaa taggtacttt cgtttgacac taggagtcta tgattacttg
     1321 gtgagcacgc aggagtttcg gtatatgaac agtcagggcc tgcttccccc aaagaactct
     1381 attgacgcgt ttaaactcaa tataaagctg ttgggcgtgg gtggaaaacc ttgcatcaag
     1441 gtggccactg tgcaggaaaa cctttacttt cagtcgtacc cctatgacgt accagattat
     1501 gcgtaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1561 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1621 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1681 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1741 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1801 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1861 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1921 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     1981 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     2041 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     2101 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     2161 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     2221 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     2281 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2761 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2881 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2941 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3001 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3061 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     3181 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     3241 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     3301 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     3361 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3421 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     3481 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     3541 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     3601 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     3661 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     3721 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     3781 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     3841 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     3901 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     3961 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     4021 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     4081 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     4141 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     4201 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     4261 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     4321 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     4381 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     4441 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     4501 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     4561 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     4621 cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg cgcctcgcca
     4681 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4741 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4801 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4861 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4921 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggagggcc
     4981 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5041 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg
     5101 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5161 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5221 gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag ataataaaat
     5281 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     5341 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     5401 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     5461 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     5521 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     5581 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5641 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5701 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5761 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5821 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5881 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5941 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6001 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6061 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6121 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     6181 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     6241 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6301 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6361 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6421 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     6481 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     6541 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     6601 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     6661 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     6721 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     6781 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     6841 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     6901 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     6961 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
