LOCUS       sequence-297364-        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"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             209..253
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    322..443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(333..352)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     387..406
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      558..604
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      634..770
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    815..886
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1112..1700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1192..1211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2200..3409
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3246..3265
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3413..4441
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4468..4516
                     /note="synthetic polyadenylation signal"
     CDS             join(4782..4850,4851..5642)
                     /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(5000..5019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5655..5791
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5943..6014
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6019..6065)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6083..6131
                     /note="synthetic polyadenylation signal"
     enhancer        6212..6515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6517..6719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6670..6690
                     /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 ttcagtcgaa agaaaccgcg gcagctaagt tcgagaggca
      241 gcacatggac agttaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt
      301 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
      361 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
      421 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
      481 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
      541 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
      601 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
      661 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
      721 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
      781 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
      841 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
      901 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
      961 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
     1021 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
     1081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1921 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     1981 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     2041 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     2101 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     2161 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg
     2221 cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca
     2281 ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     2341 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     2401 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     2461 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     2521 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     2581 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     2641 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     2701 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     2761 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     2821 gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta
     2881 ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg
     2941 gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg
     3001 cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc
     3061 ttataatgca gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag
     3121 gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg
     3181 aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa
     3241 gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt
     3301 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac
     3361 ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa
     3421 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3481 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3541 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3601 tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga
     3661 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3721 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat
     3781 cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3841 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3901 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3961 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     4021 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4081 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4141 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct
     4201 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4261 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4321 gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt
     4381 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4441 agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
     4501 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
     4561 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     4621 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     4681 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     4741 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     4801 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4861 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     4921 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     4981 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5041 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     5101 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5161 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     5221 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5281 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5341 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     5401 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5461 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5521 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5581 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5641 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     5701 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     5761 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     5821 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     5881 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     5941 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     6001 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     6061 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     6121 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     6181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     6301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     6421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     6481 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     6541 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     6601 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     6661 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     6721 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     6781 agcggccgc
//
