LOCUS       sequence-300272-        7058 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 7058)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7058)
  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-m4133424
FEATURES             Location/Qualifiers
     source          1..7058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    113..184
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(189..235)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    253..301
                     /note="synthetic polyadenylation signal"
     enhancer        382..685
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        687..889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     840..860
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1837..1857
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1861..1926
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1979..2100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1990..2009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2044..2063
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2215..2261
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2291..2427
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2472..2543
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2769..3357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2849..2868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3857..5066
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4903..4922
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5070..5669
                     /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(5070..5089)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5566..5586
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5696..5744
                     /note="synthetic polyadenylation signal"
     CDS             join(6010..6078,6079..6870)
                     /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(6228..6247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6883..7019
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
       61 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
      121 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
      181 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
      241 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
      301 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
      361 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      421 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      481 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      541 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      601 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      661 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
      721 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      781 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      841 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      901 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgcc
      961 accatgagcg ccgttaagcg gacaatcaaa gggacccatc actggttgct ccttactata
     1021 ctgacgtctt tactggtctt ggtacagagt acacaatggt cgcttttctt ttttctgtac
     1081 gagaatgcat tcctcccatt tgcgatggga atcattgcta tgtccgcctt cgctatgatg
     1141 ttcgtgaagc acaaacatgc cttcctttgc ctgtttctcc taccctcact ggcaaccgtg
     1201 gcctatttta acatggtgta catgcctgcg agctgggtca tgcgaattat gacttggctg
     1261 gacatggttg atacctccct atctggcttc aagttaaagg actgtgtgat gtatgcatca
     1321 gctgtagtgc tgctaatcct gatgacagct cgcacagtct acgatgacgg tgccaggaga
     1381 gtgtggactc tgatgaatgt cctcacctta gtttataaag tatactatgg caacgctctc
     1441 gatcaggcca ttagtatgtg ggcattgata atctccgtga cgagtaacta cagcggagtg
     1501 gtaaccactg tcatgttttt ggcacgcggg atagtgttca tgtgtgtcga atattgcccg
     1561 atctttttta tcacagggaa tactctgcaa tgcattatgc tggtttactg tttcttgggc
     1621 tatttctgca cctgttattt tggtctattc tgcctgctta ataggtactt tcgtttgaca
     1681 ctaggagtct atgattactt ggtgagcacg caggagtttc ggtatatgaa cagtcagggc
     1741 ctgcttcccc caaagaactc tattgacgcg tttaaactca atataaagct gttgggcgtg
     1801 ggtggaaaac cttgcatcaa ggtggccact gtgcaggaaa acctttactt tcagtcgatg
     1861 gattataagg accacgatgg agactacaaa gatcatgaca ttgattacaa ggacgacgat
     1921 gacaaataag ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
     1981 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2041 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2101 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2161 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
     2221 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
     2281 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     2341 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     2401 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
     2461 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
     2521 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
     2581 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
     2641 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     2701 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2761 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2821 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2881 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2941 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3001 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3061 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     3601 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     3661 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     3721 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     3781 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     3841 aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     3901 gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc
     3961 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     4021 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     4081 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     4141 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     4201 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     4261 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     4321 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     4381 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     4441 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     4501 aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac
     4561 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     4621 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     4681 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     4741 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     4801 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     4861 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     4921 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     4981 aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct
     5041 ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg gtgcgcctcg
     5101 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5161 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5221 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5281 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5341 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggagg
     5401 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5461 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggccg
     5521 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5581 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5641 ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct tagataataa
     5701 aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg
     5761 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     5821 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     5881 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     5941 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     6001 ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6061 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6121 ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6181 tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6241 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6301 aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6361 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6421 acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6481 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6541 accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt
     6601 actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca
     6661 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6721 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6781 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6841 ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa
     6901 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     6961 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     7021 atcccgaatc gtttaaactc gactctggct ctatcgaa
//
