LOCUS       sequence-301683-        6368 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 6368)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6368)
  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-m4252831
FEATURES             Location/Qualifiers
     source          1..6368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        507..1716
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1553..1572
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1720..2319
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1720..1739)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2216..2236
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2346..2394
                     /note="synthetic polyadenylation signal"
     CDS             join(2660..2728,2729..3520)
                     /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(2878..2897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3533..3669
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3821..3892
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3897..3943)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3961..4009
                     /note="synthetic polyadenylation signal"
     enhancer        4090..4393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4395..4597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4548..4568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4855..4875
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4879..4944
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4997..5118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5008..5027)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5062..5081
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5233..5279
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5309..5445
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5490..5561
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(5787..6368,1..7))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5867..5886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
       61 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      181 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      241 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
      301 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
      361 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
      421 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
      481 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
      541 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
      601 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
      661 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
      721 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
      781 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
      841 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
      901 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
      961 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     1021 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     1081 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     1141 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     1201 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     1261 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     1321 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     1381 tggggccacc agccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     1441 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     1501 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     1561 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     1621 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     1681 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     1741 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     1801 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     1861 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     1921 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     1981 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     2041 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     2101 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     2161 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     2221 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     2281 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     2341 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     2401 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     2461 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     2521 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     2581 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     2641 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     2701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2761 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2821 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     2881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2941 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     3001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3061 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     3241 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     3301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3361 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     3541 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     3601 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     3661 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     3721 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     3781 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     3841 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     3901 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     3961 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     4021 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     4081 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4141 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     4201 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     4261 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     4321 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     4381 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     4441 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     4501 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     4561 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     4621 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgccac catgtttcat
     4681 ctcgttgact ttcaggttac tatagcagag atattactaa ttattatgag gacttttaaa
     4741 gtttccattt ggaatcttga ttacatcata aacctcataa ttaaaaattt atctaagtca
     4801 ctaactgaga ataaatattc tcaattagat gaagagcaac caatggagat tgatgaaaac
     4861 ctttactttc agtcgatgga ttataaggac cacgatggag actacaaaga tcatgacatt
     4921 gattacaagg acgacgatga caaataagct agcttgactg actgagatac agcgtacctt
     4981 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     5041 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     5101 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     5161 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     5221 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     5281 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     5341 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5401 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     5461 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     5521 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     5581 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     5641 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     5701 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     5761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6361 aaaaaagg
//
