LOCUS       sequence-295965-        7372 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 7372)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7372)
  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-m4133744
FEATURES             Location/Qualifiers
     source          1..7372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             878..898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1247..1264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1341..1462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1352..1371)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1406..1425
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1577..1623
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1653..1789
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1834..1905
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2131..2719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2211..2230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3219..4428
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4265..4284
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4432..5031
                     /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(4432..4451)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4928..4948
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5058..5106
                     /note="synthetic polyadenylation signal"
     CDS             join(5372..5440,5441..6232)
                     /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(5590..5609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6245..6381
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6526..6597
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6602..6648)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6666..6714
                     /note="synthetic polyadenylation signal"
     enhancer        6795..7098
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7100..7302
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7253..7273
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc gccgttaagc ggacaatcaa agggacccat cactggttgc tccttactat
       61 actgacgtct ttactggtct tggtacagag tacacaatgg tcgcttttct tttttctgta
      121 cgagaatgca ttcctcccat ttgcgatggg aatcattgct atgtccgcct tcgctatgat
      181 gttcgtgaag cacaaacatg ccttcctttg cctgtttctc ctaccctcac tggcaaccgt
      241 ggcctatttt aacatggtgt acatgcctgc gagctgggtc atgcgaatta tgacttggct
      301 ggacatggtt gatacctccc tatctggctt caagttaaag gactgtgtga tgtatgcatc
      361 agctgtagtg ctgctaatcc tgatgacagc tcgcacagtc tacgatgacg gtgccaggag
      421 agtgtggact ctgatgaatg tcctcacctt agtttataaa gtatactatg gcaacgctct
      481 cgatcaggcc attagtatgt gggcattgat aatctccgtg acgagtaact acagcggagt
      541 ggtaaccact gtcatgtttt tggcacgcgg gatagtgttc atgtgtgtcg aatattgccc
      601 gatctttttt atcacaggga atactctgca atgcattatg ctggtttact gtttcttggg
      661 ctatttctgc acctgttatt ttggtctatt ctgcctgctt aataggtact ttcgtttgac
      721 actaggagtc tatgattact tggtgagcac gcaggagttt cggtatatga acagtcaggg
      781 cctgcttccc ccaaagaact ctattgacgc gtttaaactc aatataaagc tgttgggcgt
      841 gggtggaaaa ccttgcatca aggtggccac tgtgcaggaa aacctttact ttcagtcgat
      901 gagtgacaaa atcattcatc taaccgatga ctcctttgat acagacgtgt tgaaggccga
      961 tggtgcgata ctggtagatt tctgggctga atggtgtggc ccgtgcaaga tgatcgcacc
     1021 cattctggac gagatcgccg acgagtatca aggaaaatta actgtcgcta agcttaatat
     1081 agatcagaac ccagggacgg cacctaaata cggcatcagg ggaataccta ccctcctgct
     1141 ctttaagaat ggggaagttg ccgcgacaaa agtgggagcc ctgtctaagg gccagctaaa
     1201 agagttcttg gacgctaacc tggcaggttc aggctcgggg cacatgcacc atcaccatca
     1261 ccatagcagc gggctcgtgc cacgcggcta agctagcttg actgactgag atacagcgta
     1321 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1381 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1441 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1501 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     1561 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1621 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     1681 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     1741 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     1801 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     1861 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1921 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     1981 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     2041 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     2101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2941 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     3001 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     3061 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     3121 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     3181 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     3241 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     3301 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     3361 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     3421 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     3481 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     3541 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     3601 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     3661 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     3721 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     3781 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     3841 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     3901 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     3961 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     4021 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     4081 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     4141 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     4201 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     4261 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     4321 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     4381 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     4441 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4501 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4561 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4621 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4681 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4741 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4801 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4861 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4921 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4981 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     5041 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     5101 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     5161 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     5221 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     5281 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     5341 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     5401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5461 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     5521 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     5581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5641 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     5701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5761 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     5821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     5941 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     6001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     6241 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6301 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     6361 ttaccagaaa tttatcctta agatcgttta aactcgactc tggctctatc gaatctccgt
     6421 cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt
     6481 cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta
     6541 gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat
     6601 tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg
     6661 ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa
     6721 ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca
     6781 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     6841 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     6901 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     6961 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     7021 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     7081 gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     7141 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     7201 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     7261 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     7321 atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
