LOCUS       sequence-313667-        6317 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 6317)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6317)
  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-m4252781
FEATURES             Location/Qualifiers
     source          1..6317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             766..786
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             787..804
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    857..978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(868..887)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     922..941
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1093..1139
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1169..1305
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1350..1421
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1647..2235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1727..1746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2735..3944
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3781..3800
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3948..4547
                     /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(3948..3967)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4444..4464
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4574..4622
                     /note="synthetic polyadenylation signal"
     CDS             join(4888..4956,4957..5748)
                     /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(5106..5125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5761..5897
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6049..6120
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6125..6171)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6189..6237
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgttcca cttggtggat
      601 tttcaggtca cgatcgcaga aatactgcta attatcatgc gaaccttcaa ggttagcatt
      661 tggaacctgg actacatcat taatctcatc ataaaaaacc ttagtaagtc tttaactgag
      721 aataaatatt cccaactgga tgaggaacag ccaatggaga ttgacgaaaa cctttacttt
      781 cagtcgcacc atcaccatca ccattaagct agcttgactg actgagatac agcgtacctt
      841 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
      901 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      961 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1021 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1081 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1141 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1201 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     1261 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     1321 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     1381 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     1441 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     1501 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     1561 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     1621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2461 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     2521 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     2581 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     2641 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     2701 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     2761 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     2821 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     2881 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     2941 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     3001 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3061 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     3121 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     3181 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     3241 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     3301 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     3361 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     3421 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     3481 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     3541 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     3601 atgcagggtg gggccaccag ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     3661 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     3721 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     3781 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     3841 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta
     3901 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca
     3961 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     4021 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4081 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4141 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4201 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4261 cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4321 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4381 tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4441 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4501 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag
     4561 cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
     4621 tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     4681 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     4741 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     4801 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     4861 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     4921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4981 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     5041 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     5101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5161 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     5221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5281 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     5341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     5461 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     5521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     5761 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     5821 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     5881 cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc
     5941 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     6001 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6061 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6121 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6181 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6241 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6301 ccatatatgg agttccg
//
