LOCUS       sequence-300644-        6789 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 6789)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6789)
  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-m4252910
FEATURES             Location/Qualifiers
     source          1..6789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      249..837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     738..757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1063..1134
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1179..1315)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1345..1391)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1506..1627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1543..1562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1597..1616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1696..1740)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1741..1761)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(2019..2221)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2048..2068)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2223..2526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2607..2655
                     /note="synthetic polyadenylation signal"
     terminator      2673..2719
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2724..2795
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2947..3083)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3096..3887,3888..3956))
                     /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     3719..3738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4222..4270
                     /note="synthetic polyadenylation signal"
     CDS             complement(4297..5325)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5329..6538)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5473..5492)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ggaaatttaa atgcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
       61 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      361 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgatt
      901 taaatgaaac tcgtaatcgg agaacgggtg caatagcaag gaccagatcg aacttgactg
      961 acgcaattct gagcgtagtc gaaacccaga caagtcgatc tcgccaagta gctgacttga
     1021 ctggttaatt aagtttcgat agcccaaggt aaccaatgag attagggagg gaagaaagtt
     1081 tgatgaacta cttctgaccc ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga
     1141 tcgaaatcga gagatcgcct gcaggatagc tgacgacctt aaggataaat ttctggtaag
     1201 gaggacacgt atggaagtgg gcaagttggg gaagccgtat ccgttgctga atctggcata
     1261 tgtgggagta taagacgcgc agcgtcgcat caggcatttt tttctgcgcc aatgccggta
     1321 gatagcaatc cggcccgagg ggcacaaaaa acccctcaag acccgtttag aggccccaag
     1381 gggttatgct acgataccga tagagatggg ccaataccac atttgtagag gttttacttg
     1441 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1501 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1561 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1621 tatcttatca tgtctgtgag ctgaaggtac gctgtatctc agtcagtcaa gctagcagag
     1681 gcctgcggat ccttaactgt ccatgtgctg cctctcgaac ttagctgccg cggtttcttt
     1741 cgactgaaag taaaggtttt catcaatctc cattggttgc tcttcatcta attgagaata
     1801 tttattctca gttagtgact tagataaatt tttaattatg aggtttatga tgtaatcaag
     1861 attccaaatg gaaactttaa aagtcctcat aataattagt aatatctctg ctatagtaac
     1921 ctgaaagtca acgagatgaa acatggtggc ggccgctggc gggtgtgtcg agtggatggt
     1981 aggatcgaca aagatctgat ctgacggttc actaaaccag ctctgcttat atagacctcc
     2041 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     2101 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     2161 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     2221 agcatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     2281 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2341 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     2401 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     2461 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     2521 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactagatc
     2581 tggctacact cgatcagcag ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     2641 ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa ccccttgggg cctctaaacg
     2701 ggtcttgagg ggttttttga attgagaggt agctgaagct gctgttatga ccactagagg
     2761 gaagaagata cctgtggagc taatggtcca agatgtcggg agccgcgatc gctgccaaaa
     2821 aggtaagctg acgatagctg acgagattcg atgcccgacg agcaaatgag cgccacggct
     2881 gttcgcgtaa gctcgaaacg acggagattc gatagagcca gagtcgagtt taaacgattc
     2941 gggatcttaa ggataaattt ctggtaagga ggacacgtat ggaagtgggc aagttgggga
     3001 agccgtatcc gttgctgaat ctggcatatg tgggagtata agacgcgcag cgtcgcatca
     3061 ggcatttttt tctgcgccaa tgcacctaga atcggttacc aatgcttaat cagtgaggca
     3121 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3181 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3241 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3301 agaagtggtc ctgcaacttt atccgcctcc atccagtcta tcaactgttg ccgggaagct
     3361 agagtaagta gttcgccagt taatagttta cgcaaggttg ttgccattgc tacaggcatc
     3421 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3481 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccaatc
     3541 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3601 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgaata ctcaaccaag
     3661 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3721 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaagcg ttcttcgggg
     3781 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgcgca
     3841 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3901 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatgcga
     3961 aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa aagctaggac tcgattatac
     4021 gttgaccgat cagactatca atatctaggt ttaaacagat cggtacgctg ccgtgaccag
     4081 gcttgagtac aatatctgac gattggactt cctgggtccc aagctggatt ccgactggct
     4141 gactgccaag caacgatatg gatcgtcgag actcgcactg agaggatcgt tctcggtaca
     4201 ccaggcgcgc ctagcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     4261 cgattttatt atctaagact aggcttacat actagcttat tcctttgccc tcggacgagt
     4321 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     4381 ggcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     4441 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     4501 gagttggtca agaccaatgc ggagcatata cgcccggagc cggggcgatc ctgcaagctc
     4561 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     4621 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     4681 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     4741 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     4801 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     4861 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     4921 gccgaacccg ctcgtctggc taagatcggc ggcagcgatg gcatccattg cctccgcgac
     4981 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     5041 acggcgggag atgcaatagg tcaggctctc gctgaactcc ccaatgtcaa gcacttccgg
     5101 aatcgggagc gcggcggatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     5161 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     5221 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     5281 aaacttctcg acagacgtcg cggtgagttc aggcttttta cccatgttgt ctaacaaaaa
     5341 agccaaaaac ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat
     5401 tgacccaaat gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa
     5461 ccccgagcgc atagttcaaa accggagctt cagctactta agaagatagg tacataaaac
     5521 cgaccaaaga aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg
     5581 attcaggaga gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg
     5641 cacacctacc ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac
     5701 gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata
     5761 actgccgccc ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc
     5821 agggtcccca gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga
     5881 agggctcaag accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct
     5941 cacagcccgc ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc
     6001 gctcccccca acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg
     6061 cggtctctcc acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca
     6121 gcccagcagc ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga
     6181 cccaaatccc ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca
     6241 tcggcgttca ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc
     6301 ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc
     6361 taaaatgtcc ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc
     6421 tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca
     6481 gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag
     6541 gcgcgccccg atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt
     6601 gaacttctct cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata
     6661 ggcgctgcca agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc
     6721 gagaggtagc ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt
     6781 ggagagttc
//
