LOCUS       sequence-335176-        7503 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 7503)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7503)
  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-m4133643
FEATURES             Location/Qualifiers
     source          1..7503
                     /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             1480..1545
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1614..1735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1625..1644)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1679..1698
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1850..1896
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1926..2062
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2107..2178
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2404..2992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2484..2503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3492..4701
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4538..4557
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4705..5733
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5760..5808
                     /note="synthetic polyadenylation signal"
     CDS             join(6074..6142,6143..6934)
                     /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(6292..6311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6947..7083
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7235..7306
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7311..7357)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7375..7423
                     /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 cagtcgatgg attataagga ccacgatgga
     1501 gactacaaag atcatgacat tgattacaag gacgacgatg acaaataagg atccgcaggc
     1561 ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1621 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1681 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1741 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1801 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     1861 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1921 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     1981 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     2041 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     2101 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     2161 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     2221 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     2281 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     2341 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2401 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2461 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2521 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2581 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2641 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2701 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2761 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2821 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2881 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2941 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3001 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3061 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3121 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     3241 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3301 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     3361 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     3421 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     3481 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     3541 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     3601 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     3661 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     3721 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     3781 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     3841 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     3901 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     3961 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     4021 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     4081 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     4141 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     4201 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     4261 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     4321 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     4381 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     4441 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     4501 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     4561 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     4621 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     4681 tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc gacgtctgtc
     4741 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     4801 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     4861 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atccgccgcg
     4921 ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac ctattgcatc
     4981 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     5041 ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag ccagacgagc
     5101 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     5161 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     5221 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     5281 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     5341 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     5401 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     5461 aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg cattggtctt
     5521 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     5581 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     5641 agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     5701 cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta gtcttagata
     5761 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgctagg
     5821 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     5881 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     5941 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     6001 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     6061 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6121 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6181 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6241 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     6301 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6361 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     6421 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6481 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6541 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6601 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     6661 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     6721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6901 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     6961 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     7021 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     7081 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     7141 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     7201 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     7261 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     7321 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     7381 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     7441 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7501 ccg
//
