LOCUS       sequence-300733-        6762 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 6762)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6762)
  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-m4252886
FEATURES             Location/Qualifiers
     source          1..6762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    126..247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(163..182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     217..236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(316..333)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(334..354)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(612..814)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(641..661)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        816..1119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1200..1248
                     /note="synthetic polyadenylation signal"
     terminator      1266..1312
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1317..1388
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1540..1676)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1689..2480,2481..2549))
                     /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     2312..2331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2815..2863
                     /note="synthetic polyadenylation signal"
     CDS             complement(2890..3918)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3922..5131)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4066..4085)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5631..6219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6120..6139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6445..6516
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6561..6697)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(join(6727..6762,1..11))
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gggttatgct acgataccga tagagatggg ccaataccac atttgtagag gttttacttg
       61 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
      121 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
      181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      241 tatcttatca tgtctgtgag ctgaaggtac gctgtatctc agtcagtcaa gctagcagag
      301 gcctgcggat ccttaatggt gatggtgatg gtgcgactga aagtaaaggt tttcgtcaat
      361 ctccattggc tgttcctcat ccagttggga atatttattc tcagttaaag acttactaag
      421 gttttttatg atgagattaa tgatgtagtc caggttccaa atgctaacct tgaaggttcg
      481 catgataatt agcagtattt ctgcgatcgt gacctgaaaa tccaccaagt ggaacatggt
      541 ggcggccgct ggcgggtgtg tcgagtggat ggtaggatcg acaaagatct gatctgacgg
      601 ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
      661 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
      721 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
      781 ccacgcccat tgatgtactg ccaaaaccgc atcagcatgg taatagcgat gactaatacg
      841 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
      901 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
      961 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     1021 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     1081 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     1141 tgaactaatg accccgtaat tgattactag atctggctac actcgatcag cagttagata
     1201 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgcaacg
     1261 attgatagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaattgaga
     1321 ggtagctgaa gctgctgtta tgaccactag agggaagaag atacctgtgg agctaatggt
     1381 ccaagatgtc gggagccgcg atcgctgcca aaaaggtaag ctgacgatag ctgacgagat
     1441 tcgatgcccg acgagcaaat gagcgccacg gctgttcgcg taagctcgaa acgacggaga
     1501 ttcgatagag ccagagtcga gtttaaacga ttcgggatct taaggataaa tttctggtaa
     1561 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
     1621 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgcacct
     1681 agaatcggtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1741 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1801 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1861 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1921 tccatccagt ctatcaactg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1981 ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2041 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2101 aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca gaagtaagtt ggccgcagtg
     2161 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2221 tgcttttctg tgactggtga atactcaacc aagtcattct gagaatagtg tatgcggcga
     2281 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2341 aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2401 ttgagatcca gttcgatgta acccactcgc gcacccaact gatcttcagc atcttttact
     2461 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2521 agggcgacac ggaaatgttg aatactcatg cgaaagcctc ctgctgatcc tgtctcttga
     2581 tagatgtttg caaaagctag gactcgatta tacgttgacc gatcagacta tcaatatcta
     2641 ggtttaaaca gatcggtacg ctgccgtgac caggcttgag tacaatatct gacgattgga
     2701 cttcctgggt cccaagctgg attccgactg gctgactgcc aagcaacgat atggatcgtc
     2761 gagactcgca ctgagaggat cgttctcggt acaccaggcg cgcctagcgt tacacacaca
     2821 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaag actaggctta
     2881 catactagct tattcctttg ccctcggacg agtgctgggg cgtcggtttc cactatcggc
     2941 gagtacttct acacagccat cggtccagac ggcggcgctt ctgcgggcga tttgtgtacg
     3001 cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc
     3061 atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat
     3121 atacgcccgg agccggggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt
     3181 ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg
     3241 ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc cattgtccgt
     3301 caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc
     3361 ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg tgtcgtccat
     3421 cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat cacgccatgt
     3481 agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc
     3541 ggcggcagcg atggcatcca ttgcctccgc gaccggctgc agaacagcgg gcagttcggt
     3601 ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat aggtcaggct
     3661 ctcgctgaac tccccaatgt caagcacttc cggaatcggg agcgcggcgg atgcaaagtg
     3721 ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc gcaggacata
     3781 tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg agagctgcat
     3841 caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag
     3901 ttcaggcttt ttacccatgt tgtctaacaa aaaagccaaa aacggccaga atttagcgga
     3961 caatttacaa gtctaacact gaaaattaca tattgaccca aatgattaca tttcaaaagg
     4021 tgcctaaaaa acttcacaaa acacactcgc caaccccgag cgcatagttc aaaaccggag
     4081 cttcagctac ttaagaagat aggtacataa aaccgaccaa agaaactgac gcctcactta
     4141 tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc ctaggcccga
     4201 accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggctggt ggccccaccc
     4261 tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg cgctcccaaa
     4321 ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac agacgacaaa
     4381 ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg cccagcccat
     4441 ctcacccgaa taagagcttt cccgcattag cgaagggctc aagaccttgg gttcttgccg
     4501 cccaccatgc cccccacctt gtttcaacga cctcacagcc cgcctcacaa gcgtcttcca
     4561 ttcaagactc gggaacagcc gccattttgc tgcgctcccc ccaaccccca gttcagggca
     4621 accttgctcg cggacccaga ctacagccct tggcggtctc tccacacgct tccgtcccac
     4681 cgagcggccc ggcggccacg aaagccccgg ccagcccagc agcccgctac tcaccaagtg
     4741 acgatcacag cgatccacaa acaagaaccg cgacccaaat cccggctgcg acggaactag
     4801 ctgtgccaca cccggcgcgt ccttatataa tcatcggcgt tcaccgcccc acggagatcc
     4861 ctccgcagaa tcgccgagaa gggactactt ttcctcgcct gttccgctct ctggaaagaa
     4921 aaccagtgcc ctagagtcac ccaagtcccg tcctaaaatg tccttctgct gatactgggg
     4981 ttctaaggcc gagtcttatg agcagcgggc cgctgtcctg agcgtccggg cggaaggatc
     5041 aggacgctcc tgcgcccttc gtctgacgtg gcagcgctcg ccgtgacgag gggggcgccc
     5101 gcgggaggcg ccaaaacccg gcgcggaggc caggcgcgcc ccgatttcga tattcctcgg
     5161 atctcggata gatcgggatt gaggatgtag gttgaacttc tctcgggtga tcccgatctc
     5221 ggatattcgg gattggagta atacctggcg ataggcgctg ccaagcaata gccaaggcgc
     5281 aaggccggcc aagagacttg cgatagttcg ttcgagaggt agcctgcaag atggatcgga
     5341 cgaaaggttt gaagattttc cgacgagggc cttggagagt tcggaaattt aaatgcaact
     5401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6241 ttacggttcc tggccttttg ctggcctttt gctcacatga tttaaatgaa actcgtaatc
     6301 ggagaacggg tgcaatagca aggaccagat cgaacttgac tgacgcaatt ctgagcgtag
     6361 tcgaaaccca gacaagtcga tctcgccaag tagctgactt gactggttaa ttaagtttcg
     6421 atagcccaag gtaaccaatg agattaggga gggaagaaag tttgatgaac tacttctgac
     6481 ccctagtggt gtccagaaaa gaccattaaa ggaatgtctt gatcgaaatc gagagatcgc
     6541 ctgcaggata gctgacgacc ttaaggataa atttctggta aggaggacac gtatggaagt
     6601 gggcaagttg gggaagccgt atccgttgct gaatctggca tatgtgggag tataagacgc
     6661 gcagcgtcgc atcaggcatt tttttctgcg ccaatgccgg tagatagcaa tccggcccga
     6721 ggggcacaaa aaacccctca agacccgttt agaggcccca ag
//
