LOCUS       sequence-307317-        6377 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 6377)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6377)
  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-m4252820
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      66..112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    117..188
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(340..476)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(489..1280,1281..1349))
                     /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     1112..1131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1615..1663
                     /note="synthetic polyadenylation signal"
     CDS             complement(1690..2289)
                     /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(1773..1793)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2270..2289
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2293..3502)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2437..2456)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4002..4590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4491..4510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4816..4887
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4932..5068)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5098..5144)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5259..5380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5296..5315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5350..5369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5433..5510)
                     /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             complement(5511..5531)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5789..5991)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5818..5838)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5993..6296
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6377,1..48)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgcaacg
       61 attgatagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaattgaga
      121 ggtagctgaa gctgctgtta tgaccactag agggaagaag atacctgtgg agctaatggt
      181 ccaagatgtc gggagccgcg atcgctgcca aaaaggtaag ctgacgatag ctgacgagat
      241 tcgatgcccg acgagcaaat gagcgccacg gctgttcgcg taagctcgaa acgacggaga
      301 ttcgatagag ccagagtcga gtttaaacga ttcgggatct taaggataaa tttctggtaa
      361 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
      421 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgcacct
      481 agaatcggtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
      541 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
      601 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
      661 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
      721 tccatccagt ctatcaactg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
      781 ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
      841 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
      901 aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca gaagtaagtt ggccgcagtg
      961 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1021 tgcttttctg tgactggtga atactcaacc aagtcattct gagaatagtg tatgcggcga
     1081 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     1141 aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1201 ttgagatcca gttcgatgta acccactcgc gcacccaact gatcttcagc atcttttact
     1261 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     1321 agggcgacac ggaaatgttg aatactcatg cgaaagcctc ctgctgatcc tgtctcttga
     1381 tagatgtttg caaaagctag gactcgatta tacgttgacc gatcagacta tcaatatcta
     1441 ggtttaaaca gatcggtacg ctgccgtgac caggcttgag tacaatatct gacgattgga
     1501 cttcctgggt cccaagctgg attccgactg gctgactgcc aagcaacgat atggatcgtc
     1561 gagactcgca ctgagaggat cgttctcggt acaccaggcg cgcctagcgt tacacacaca
     1621 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaag actaggctta
     1681 catactagcc taggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac
     1741 ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc
     1801 ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcg gcctccactc cggggagcac
     1861 gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa
     1921 ccacgcgggc tccttgggcc ggtgcggcgc caggaggccc tccatctgtt gctgcgcggc
     1981 cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc
     2041 ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac
     2101 cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc
     2161 gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc
     2221 gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta
     2281 ctcggtcatg ttgtctaaca aaaaagccaa aaacggccag aatttagcgg acaatttaca
     2341 agtctaacac tgaaaattac atattgaccc aaatgattac atttcaaaag gtgcctaaaa
     2401 aacttcacaa aacacactcg ccaaccccga gcgcatagtt caaaaccgga gcttcagcta
     2461 cttaagaaga taggtacata aaaccgacca aagaaactga cgcctcactt atccctcccc
     2521 tcaccagagg tccggcgcct gtcgattcag gagagcctac cctaggcccg aaccctgcgt
     2581 cctgcgacgg agaaaagcct accgcacacc taccggctgg tggccccacc ctgcattata
     2641 agccaacaga acgggtgacg tcacgacacg acgagggcgc gcgctcccaa aggtacgggt
     2701 gcactgccca acggcaccgc cataactgcc gcccccgcaa cagacgacaa accgagttct
     2761 ccagtcagtg acaaacttca cgtcagggtc cccagatggt gcccagccca tctcacccga
     2821 ataagagctt tcccgcatta gcgaagggct caagaccttg ggttcttgcc gcccaccatg
     2881 ccccccacct tgtttcaacg acctcacagc ccgcctcaca agcgtcttcc attcaagact
     2941 cgggaacagc cgccattttg ctgcgctccc cccaaccccc agttcagggc aaccttgctc
     3001 gcggacccag actacagccc ttggcggtct ctccacacgc ttccgtccca ccgagcggcc
     3061 cggcggccac gaaagccccg gccagcccag cagcccgcta ctcaccaagt gacgatcaca
     3121 gcgatccaca aacaagaacc gcgacccaaa tcccggctgc gacggaacta gctgtgccac
     3181 acccggcgcg tccttatata atcatcggcg ttcaccgccc cacggagatc cctccgcaga
     3241 atcgccgaga agggactact tttcctcgcc tgttccgctc tctggaaaga aaaccagtgc
     3301 cctagagtca cccaagtccc gtcctaaaat gtccttctgc tgatactggg gttctaaggc
     3361 cgagtcttat gagcagcggg ccgctgtcct gagcgtccgg gcggaaggat caggacgctc
     3421 ctgcgccctt cgtctgacgt ggcagcgctc gccgtgacga ggggggcgcc cgcgggaggc
     3481 gccaaaaccc ggcgcggagg ccaggcgcgc cccgatttcg atattcctcg gatctcggat
     3541 agatcgggat tgaggatgta ggttgaactt ctctcgggtg atcccgatct cggatattcg
     3601 ggattggagt aatacctggc gataggcgct gccaagcaat agccaaggcg caaggccggc
     3661 caagagactt gcgatagttc gttcgagagg tagcctgcaa gatggatcgg acgaaaggtt
     3721 tgaagatttt ccgacgaggg ccttggagag ttcggaaatt taaatgcaac tatggatgaa
     3781 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3841 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3901 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3961 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4021 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4081 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4141 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4201 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4261 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4321 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4381 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4441 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4501 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4561 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4621 ctggcctttt gctggccttt tgctcacatg atttaaatga aactcgtaat cggagaacgg
     4681 gtgcaatagc aaggaccaga tcgaacttga ctgacgcaat tctgagcgta gtcgaaaccc
     4741 agacaagtcg atctcgccaa gtagctgact tgactggtta attaagtttc gatagcccaa
     4801 ggtaaccaat gagattaggg agggaagaaa gtttgatgaa ctacttctga cccctagtgg
     4861 tgtccagaaa agaccattaa aggaatgtct tgatcgaaat cgagagatcg cctgcaggat
     4921 agctgacgac cttaaggata aatttctggt aaggaggaca cgtatggaag tgggcaagtt
     4981 ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg cgcagcgtcg
     5041 catcaggcat ttttttctgc gccaatgccg gtagatagca atccggcccg aggggcacaa
     5101 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctacgatac cgatagagat
     5161 gggccaatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5221 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     5281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt gagctgaagg
     5401 tacgctgtat ctcagtcagt caagctagct tagagggcac cagacgagga tatctttttg
     5461 aatctgttgg ctgcactcac cgcgataaaa ttctttttcc atcgcctctt cgactgaaag
     5521 taaaggtttt catcaatctc cattggttgc tcttcatcta attgagaata tttattctca
     5581 gttagtgact tagataaatt tttaattatg aggtttatga tgtaatcaag attccaaatg
     5641 gaaactttaa aagtcctcat aataattagt aatatctctg ctatagtaac ctgaaagtca
     5701 acgagatgaa acatggtggc ggccgctggc gggtgtgtcg agtggatggt aggatcgaca
     5761 aagatctgat ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac
     5821 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     5881 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     5941 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc agcatggtaa
     6001 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     6061 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     6121 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     6181 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     6241 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     6301 actccatata tgggctatga actaatgacc ccgtaattga ttactagatc tggctacact
     6361 cgatcagcag ttagata
//
