LOCUS       sequence-297445-        6344 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 6344)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6344)
  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-m4252829
FEATURES             Location/Qualifiers
     source          1..6344
                     /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    306..427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(317..336)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     371..390
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      542..588
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      618..754
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    799..870
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1096..1684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1176..1195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2184..3393
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3230..3249
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3397..3996
                     /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(3397..3416)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3893..3913
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4023..4071
                     /note="synthetic polyadenylation signal"
     CDS             join(4337..4405,4406..5197)
                     /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(4555..4574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5210..5346
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5498..5569
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5574..5620)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5638..5686
                     /note="synthetic polyadenylation signal"
     enhancer        5767..6070
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6072..6274
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6225..6245
                     /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 agttaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
      301 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      361 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      421 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      481 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
      541 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
      601 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
      661 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
      721 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
      781 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
      841 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
      901 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
      961 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     1021 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1921 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     1981 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     2041 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     2101 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     2161 aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc
     2221 ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct
     2281 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     2341 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     2401 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     2461 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     2521 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     2581 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga
     2641 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct
     2701 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag
     2761 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa
     2821 gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc
     2881 tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     2941 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     3001 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     3061 ggccaccagc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     3121 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     3181 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     3241 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     3301 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct
     3361 aaattctggc cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg
     3421 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     3481 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     3541 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     3601 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     3661 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     3721 atggagggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     3781 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     3841 gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     3901 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     3961 cgcacctggt gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag
     4021 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct
     4081 aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     4141 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     4201 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     4261 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     4321 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4381 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4441 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4501 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     4561 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4621 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     4681 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4741 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4801 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4861 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     4921 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     4981 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5041 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5101 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5161 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     5221 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     5281 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     5341 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     5401 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     5461 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     5521 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     5581 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     5641 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     5701 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5761 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     5821 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     5881 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5941 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     6001 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6061 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     6121 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     6181 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     6241 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     6301 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
