LOCUS       sequence-295920-        7439 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 7439)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7439)
  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-m4133456
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             872..892
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             893..910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    979..1100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(990..1009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1044..1063
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1215..1261
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1291..1427
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1472..1543
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1769..2357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1849..1868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2857..4066
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3903..3922
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4070..5098
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5125..5173
                     /note="synthetic polyadenylation signal"
     CDS             join(5439..5507,5508..6299)
                     /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(5657..5676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6312..6448
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6593..6664
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6669..6715)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6733..6781
                     /note="synthetic polyadenylation signal"
     enhancer        6862..7165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7167..7369
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7320..7340
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7438..7439,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gtgaaaagaa caatcaaggg tacacaccac tggttgttac tcacaatttt
       61 gacttcactt ttagttttag tccagagtac tcaatggtct ttgttctttt ttttgtatga
      121 aaatgccttt ttaccttttg ctatgggtat tattgctatg tctgcttttg caatgatgtt
      181 tgtcaaacat aagcatgcat ttctctgttt gtttttgtta ccttctcttg ccactgtagc
      241 ttattttaat atggtctata tgcctgctag ttgggtgatg cgtattatga catggttgga
      301 tatggttgat actagtttgt ctggttttaa gctaaaagac tgtgttatgt atgcatcagc
      361 tgtagtgtta ctaatcctta tgacagcaag aactgtgtat gatgatggtg ctaggagagt
      421 gtggacactt atgaatgtct tgacactcgt ttataaagtt tattatggta atgctttaga
      481 tcaagccatt tccatgtggg ctcttataat ctctgttact tctaactact caggtgtagt
      541 tacaactgtc atgtttttgg ccagaggtat tgtttttatg tgtgttgagt attgccctat
      601 tttcttcata actggtaata cacttcagtg tataatgcta gtttattgtt tcttaggcta
      661 tttttgtact tgttactttg gcctcttttg tttactcaac cgctacttta gactgactct
      721 tggtgtttat gattacttag tttctacaca ggagtttaga tatatgaatt cacagggact
      781 actcccaccc aagaatagca tagatgcctt caaactcaac attaaattgt tgggtgttgg
      841 tggcaaacct tgtatcaaag tagccactgt agaaaacctt tactttcagt cgcaccatca
      901 ccatcaccat taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
      961 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1021 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1081 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1141 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
     1201 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1261 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
     1321 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     1381 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     1441 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     1501 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     1561 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     1621 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     1681 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     1741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2581 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     2641 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     2701 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     2761 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     2821 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     2881 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     2941 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     3001 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     3061 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     3121 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     3181 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     3241 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     3301 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     3361 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     3421 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     3481 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     3541 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     3601 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     3661 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     3721 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     3781 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     3841 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     3901 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     3961 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     4021 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     4081 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4141 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4201 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4261 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     4321 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4381 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     4441 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4501 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4561 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4621 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4681 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4741 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4801 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     4861 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4921 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4981 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     5041 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     5101 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     5161 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     5221 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     5281 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     5341 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     5401 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     5461 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5521 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     5581 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     5641 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5701 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     5761 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5821 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     5881 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5941 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     6001 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     6061 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6121 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6181 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6241 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6301 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     6361 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6421 gtcctcctta ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa
     6481 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6541 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     6601 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     6661 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     6721 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     6781 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     6841 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6901 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6961 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7021 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7081 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     7141 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     7201 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     7261 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     7321 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     7381 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
