LOCUS       sequence-297446-        7299 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 7299)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7299)
  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-m4252933
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             242..319
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             347..367
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    832..953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(843..862)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     897..916
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1068..1114
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1144..1280
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1325..1396
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1622..2210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1702..1721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2710..3919
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3756..3775
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3923..4951
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4978..5026
                     /note="synthetic polyadenylation signal"
     CDS             join(5292..5360,5361..6152)
                     /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(5510..5529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6165..6301
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6453..6524
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6529..6575)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6593..6641
                     /note="synthetic polyadenylation signal"
     enhancer        6722..7025
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7027..7229
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7180..7200
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttt catctcgttg actttcaggt tactatagca gagatattac taattattat
       61 gaggactttt aaagtttcca tttggaatct tgattacatc ataaacctca taattaaaaa
      121 tttatctaag tcactaactg agaataaata ttctcaatta gatgaagagc aaccaatgga
      181 gattgatgaa aacctttact ttcagtcggg acggcgaatc ccggggctga taaatccctg
      241 gaaaaggaga tggaagaaga actttattgc agtgtctgcg gctaaccggt tcaaaaagat
      301 cagttcaagc ggcgccctag attacgacat tcctacgacc gcctccgaga atctctattt
      361 ccagggtgaa tttggcttgg cacaacatga cgaggccgtc gataacaagt ttaataaaga
      421 acagcagaac gctttctatg agatccttca cttaccaaat ctgaacgaag agcaacgcaa
      481 cgctttcatt cagagcctga aagacgatcc ctcgcagtct gcaaatctcc tggccgaagc
      541 caagaagctg aatgacgcac aggctccaaa agttgataac aagtttaaca aggagcagca
      601 gaatgctttc tacgagatat tgcaccttcc taatctgaat gaggaacaaa gaaatgcatt
      661 tatccagtcc ctaaaagacg atccatcaca gagtgccaac ctgcttgcgg aagccaaaaa
      721 gctcaatgat gcccaggccc cgaaagtaga cgcgaaccat caataaggat ccgcaggcct
      781 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
      841 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      901 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      961 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1021 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     1081 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     1141 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     1201 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     1261 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     1321 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     1381 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     1441 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     1501 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     1561 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2041 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2281 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2341 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     2461 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     2521 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     2581 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     2641 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     2701 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     2761 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     2821 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2881 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2941 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     3001 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     3061 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     3121 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     3181 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3241 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3301 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3361 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     3421 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     3481 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     3541 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt
     3601 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     3661 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     3721 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     3781 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     3841 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     3901 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     3961 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     4021 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     4081 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     4141 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     4201 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4261 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     4321 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4381 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4441 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4501 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4561 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4621 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4681 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     4741 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4801 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4861 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4921 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     4981 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     5041 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     5101 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     5161 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     5221 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     5281 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5341 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5401 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5461 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5521 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5581 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5641 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5701 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5761 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5821 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     5881 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     5941 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6001 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6061 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6121 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     6181 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     6241 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     6301 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     6361 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     6421 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     6481 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     6541 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     6601 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     6661 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6721 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6781 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6841 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6901 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6961 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     7021 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     7081 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     7141 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     7201 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     7261 cgatcctacc atccactcga cacacccgcc agcggccgc
//
