LOCUS       sequence-299531-        4437 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 4437)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4437)
  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-m4133582
FEATURES             Location/Qualifiers
     source          1..4437
                     /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"
     CDS             join(2430..2498,2499..3290)
                     /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(2648..2667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3303..3439
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3591..3662
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3667..3713)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3731..3779
                     /note="synthetic polyadenylation signal"
     enhancer        3860..4163
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4165..4367
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4318..4338
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc cacttggtgg attttcaggt cacgatcgca gaaatactgc taattatcat
       61 gcgaaccttc aaggttagca tttggaacct ggactacatc attaatctca tcataaaaaa
      121 ccttagtaag tctttaactg agaataaata ttcccaactg gatgaggaac agccaatgga
      181 gattgacgaa 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 cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     2221 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     2281 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     2341 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     2401 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     2461 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2521 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     2581 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     2641 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2701 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     2761 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2821 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     2881 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2941 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     3001 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     3061 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3121 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3181 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3241 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     3301 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     3361 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     3421 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     3481 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     3541 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     3601 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     3661 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     3721 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     3781 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     3841 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3901 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3961 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4021 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4081 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4141 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     4201 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4261 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4321 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     4381 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
