LOCUS       sequence-300692-        6326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6326)
  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-m4252778
FEATURES             Location/Qualifiers
     source          1..6326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      42..88
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    93..164
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(316..452)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(465..1256,1257..1325))
                     /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     1088..1107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1591..1639
                     /note="synthetic polyadenylation signal"
     CDS             complement(1666..2265)
                     /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(1749..1769)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2246..2265
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2269..3478)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2413..2432)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      3978..4566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4467..4486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4792..4863
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4908..5044)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5074..5120)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5235..5356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5272..5291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5326..5345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5409..5435)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5436..5456)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5714..5916)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5743..5763)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5918..6221
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6302..6326,1..24)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ggatgtgtgt tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc
       61 ctctaaacgg gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac
      121 cactagaggg aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg
      181 ctgccaaaaa ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc
      241 gccacggctg ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt
      301 aaacgattcg ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca
      361 agttggggaa gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc
      421 gtcgcatcag gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc
      481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
      601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
      661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc
      721 cgggaagcta gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct
      781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      901 cctccaatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
      961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac
     1021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt
     1141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1201 actcgcgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1321 ctcatgcgaa agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact
     1381 cgattatacg ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc
     1441 cgtgaccagg cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc
     1501 cgactggctg actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt
     1561 ctcggtacac caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc
     1621 gatggatagc gattttatta tctaagacta ggcttacata ctagcctagg caccgggctt
     1681 gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa
     1741 gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac
     1801 cccggcgcgc tcggcggcct ccactccggg gagcacgacg gcgctgccca gacccttgcc
     1861 ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg
     1921 cggcgccagg aggccctcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc
     1981 catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca
     2041 gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg
     2101 cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt
     2161 gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac
     2221 gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatgttgt ctaacaaaaa
     2281 agccaaaaac ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat
     2341 tgacccaaat gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa
     2401 ccccgagcgc atagttcaaa accggagctt cagctactta agaagatagg tacataaaac
     2461 cgaccaaaga aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg
     2521 attcaggaga gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg
     2581 cacacctacc ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac
     2641 gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata
     2701 actgccgccc ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc
     2761 agggtcccca gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga
     2821 agggctcaag accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct
     2881 cacagcccgc ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc
     2941 gctcccccca acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg
     3001 cggtctctcc acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca
     3061 gcccagcagc ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga
     3121 cccaaatccc ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca
     3181 tcggcgttca ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc
     3241 ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc
     3301 taaaatgtcc ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc
     3361 tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca
     3421 gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag
     3481 gcgcgccccg atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt
     3541 gaacttctct cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata
     3601 ggcgctgcca agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc
     3661 gagaggtagc ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt
     3721 ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa atagacagat cgctgagata
     3781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4621 cacatgattt aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg accagatcga
     4681 acttgactga cgcaattctg agcgtagtcg aaacccagac aagtcgatct cgccaagtag
     4741 ctgacttgac tggttaatta agtttcgata gcccaaggta accaatgaga ttagggaggg
     4801 aagaaagttt gatgaactac ttctgacccc tagtggtgtc cagaaaagac cattaaagga
     4861 atgtcttgat cgaaatcgag agatcgcctg caggatagct gacgacctta aggataaatt
     4921 tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa
     4981 tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca
     5041 atgccggtag atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga
     5101 ggccccaagg ggttatgcta cgataccgat agagatgggc caataccaca tttgtagagg
     5161 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5221 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5281 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5341 tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag
     5401 ctagcttacg cataatctgg tacgtcatag gggtacgact gaaagtaaag gttttcatca
     5461 atctccattg gttgctcttc atctaattga gaatatttat tctcagttag tgacttagat
     5521 aaatttttaa ttatgaggtt tatgatgtaa tcaagattcc aaatggaaac tttaaaagtc
     5581 ctcataataa ttagtaatat ctctgctata gtaacctgaa agtcaacgag atgaaacatg
     5641 gtggcggccg ctggcgggtg tgtcgagtgg atggtaggat cgacaaagat ctgatctgac
     5701 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     5761 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     5821 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     5881 atccacgccc attgatgtac tgccaaaacc gcatcagcat ggtaatagcg atgactaata
     5941 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     6001 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     6061 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     6121 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     6181 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     6241 tatgaactaa tgaccccgta attgattact agatctggct acactcgatc agcagttaga
     6301 taataaaatc gctatccatc gaagat
//
