LOCUS       sequence-300717-        6813 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 6813)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6813)
  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-m4252925
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      263..851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     752..771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1077..1148
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1193..1329)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1359..1405)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1520..1641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1557..1576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1611..1630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1710..1775)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1779..1799)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(2057..2259)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2086..2106)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2261..2564
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2645..2693
                     /note="synthetic polyadenylation signal"
     terminator      2711..2757
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2762..2833
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2985..3121)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3134..3925,3926..3994))
                     /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     3757..3776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4260..4308
                     /note="synthetic polyadenylation signal"
     CDS             complement(4335..5363)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5367..6576)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5511..5530)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gccttggaga gttcggaaat ttaaatgcaa ctatggatga acgaaataga cagatcgctg
       61 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
      121 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
      181 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
      241 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      301 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      361 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      421 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      481 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      541 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      601 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      661 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      721 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      781 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      841 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      901 ttgctcacat gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag caaggaccag
      961 atcgaacttg actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca
     1021 agtagctgac ttgactggtt aattaagttt cgatagccca aggtaaccaa tgagattagg
     1081 gagggaagaa agtttgatga actacttctg acccctagtg gtgtccagaa aagaccatta
     1141 aaggaatgtc ttgatcgaaa tcgagagatc gcctgcagga tagctgacga ccttaaggat
     1201 aaatttctgg taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg
     1261 ctgaatctgg catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg
     1321 cgccaatgcc ggtagatagc aatccggccc gaggggcaca aaaaacccct caagacccgt
     1381 ttagaggccc caaggggtta tgctacgata ccgatagaga tgggccaata ccacatttgt
     1441 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1501 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1621 caaactcatc aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag
     1681 tcaagctagc agaggcctgc ggatccttat ttgtcatcgt cgtccttgta atcaatgtca
     1741 tgatctttgt agtctccatc gtggtcctta taatccatcg actgaaagta aaggttttcg
     1801 tcaatctcca ttggctgttc ctcatccagt tgggaatatt tattctcagt taaagactta
     1861 ctaaggtttt ttatgatgag attaatgatg tagtccaggt tccaaatgct aaccttgaag
     1921 gttcgcatga taattagcag tatttctgcg atcgtgacct gaaaatccac caagtggaac
     1981 atggtggcgg ccgctggcgg gtgtgtcgag tggatggtag gatcgacaaa gatctgatct
     2041 gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     2101 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     2161 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     2221 gctatccacg cccattgatg tactgccaaa accgcatcag catggtaata gcgatgacta
     2281 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     2341 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     2401 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     2461 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     2521 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     2581 ggctatgaac taatgacccc gtaattgatt actagatctg gctacactcg atcagcagtt
     2641 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     2701 caacgattga tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgaat
     2761 tgagaggtag ctgaagctgc tgttatgacc actagaggga agaagatacc tgtggagcta
     2821 atggtccaag atgtcgggag ccgcgatcgc tgccaaaaag gtaagctgac gatagctgac
     2881 gagattcgat gcccgacgag caaatgagcg ccacggctgt tcgcgtaagc tcgaaacgac
     2941 ggagattcga tagagccaga gtcgagttta aacgattcgg gatcttaagg ataaatttct
     3001 ggtaaggagg acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct
     3061 ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg
     3121 cacctagaat cggttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3361 ccgcctccat ccagtctatc aactgttgcc gggaagctag agtaagtagt tcgccagtta
     3421 atagtttacg caaggttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3541 tgtgcaaaaa agcggttagc tccttcggtc ctccaatcgt tgtcagaagt aagttggccg
     3601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3661 taagatgctt ttctgtgact ggtgaatact caaccaagtc attctgagaa tagtgtatgc
     3721 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3781 ctttaaaagt gctcatcatt ggaaagcgtt cttcggggcg aaaactctca aggatcttac
     3841 cgctgttgag atccagttcg atgtaaccca ctcgcgcacc caactgatct tcagcatctt
     3901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3961 gaataagggc gacacggaaa tgttgaatac tcatgcgaaa gcctcctgct gatcctgtct
     4021 cttgatagat gtttgcaaaa gctaggactc gattatacgt tgaccgatca gactatcaat
     4081 atctaggttt aaacagatcg gtacgctgcc gtgaccaggc ttgagtacaa tatctgacga
     4141 ttggacttcc tgggtcccaa gctggattcc gactggctga ctgccaagca acgatatgga
     4201 tcgtcgagac tcgcactgag aggatcgttc tcggtacacc aggcgcgcct agcgttacac
     4261 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaagactag
     4321 gcttacatac tagcttattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta
     4381 tcggcgagta cttctacaca gccatcggtc cagacggcgg cgcttctgcg ggcgatttgt
     4441 gtacgcccga cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa
     4501 gctgcatcat cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg
     4561 agcatatacg cccggagccg gggcgatcct gcaagctccg gatgcctccg ctcgaagtag
     4621 cgcgtctgct gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg
     4681 tattgggaat ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg
     4741 tccgtcagga cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag
     4801 tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg
     4861 tccatcacag tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc
     4921 catgtagtgt attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta
     4981 agatcggcgg cagcgatggc atccattgcc tccgcgaccg gctgcagaac agcgggcagt
     5041 tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc
     5101 aggctctcgc tgaactcccc aatgtcaagc acttccggaa tcgggagcgc ggcggatgca
     5161 aagtgccgat aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg
     5221 acatatccac gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc
     5281 tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg
     5341 gtgagttcag gctttttacc catgttgtct aacaaaaaag ccaaaaacgg ccagaattta
     5401 gcggacaatt tacaagtcta acactgaaaa ttacatattg acccaaatga ttacatttca
     5461 aaaggtgcct aaaaaacttc acaaaacaca ctcgccaacc ccgagcgcat agttcaaaac
     5521 cggagcttca gctacttaag aagataggta cataaaaccg accaaagaaa ctgacgcctc
     5581 acttatccct cccctcacca gaggtccggc gcctgtcgat tcaggagagc ctaccctagg
     5641 cccgaaccct gcgtcctgcg acggagaaaa gcctaccgca cacctaccgg ctggtggccc
     5701 caccctgcat tataagccaa cagaacgggt gacgtcacga cacgacgagg gcgcgcgctc
     5761 ccaaaggtac gggtgcactg cccaacggca ccgccataac tgccgccccc gcaacagacg
     5821 acaaaccgag ttctccagtc agtgacaaac ttcacgtcag ggtccccaga tggtgcccag
     5881 cccatctcac ccgaataaga gctttcccgc attagcgaag ggctcaagac cttgggttct
     5941 tgccgcccac catgcccccc accttgtttc aacgacctca cagcccgcct cacaagcgtc
     6001 ttccattcaa gactcgggaa cagccgccat tttgctgcgc tccccccaac ccccagttca
     6061 gggcaacctt gctcgcggac ccagactaca gcccttggcg gtctctccac acgcttccgt
     6121 cccaccgagc ggcccggcgg ccacgaaagc cccggccagc ccagcagccc gctactcacc
     6181 aagtgacgat cacagcgatc cacaaacaag aaccgcgacc caaatcccgg ctgcgacgga
     6241 actagctgtg ccacacccgg cgcgtcctta tataatcatc ggcgttcacc gccccacgga
     6301 gatccctccg cagaatcgcc gagaagggac tacttttcct cgcctgttcc gctctctgga
     6361 aagaaaacca gtgccctaga gtcacccaag tcccgtccta aaatgtcctt ctgctgatac
     6421 tggggttcta aggccgagtc ttatgagcag cgggccgctg tcctgagcgt ccgggcggaa
     6481 ggatcaggac gctcctgcgc ccttcgtctg acgtggcagc gctcgccgtg acgagggggg
     6541 cgcccgcggg aggcgccaaa acccggcgcg gaggccaggc gcgccccgat ttcgatattc
     6601 ctcggatctc ggatagatcg ggattgagga tgtaggttga acttctctcg ggtgatcccg
     6661 atctcggata ttcgggattg gagtaatacc tggcgatagg cgctgccaag caatagccaa
     6721 ggcgcaaggc cggccaagag acttgcgata gttcgttcga gaggtagcct gcaagatgga
     6781 tcggacgaaa ggtttgaaga ttttccgacg agg
//
