LOCUS       sequence-297353-        4461 bp    DNA     circular SYN 15-APR-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 4461)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4461)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133493
FEATURES             Location/Qualifiers
     source          1..4461
                     /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             212..277
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    330..451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(341..360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     395..414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      566..612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      642..778
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    823..894
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1120..1708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1200..1219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2454..2522,2523..3314)
                     /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(2672..2691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3327..3463
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3615..3686
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3691..3737)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3755..3803
                     /note="synthetic polyadenylation signal"
     enhancer        3884..4187
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4189..4391
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4342..4362
                     /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 ttcagtcgat ggattataag gaccacgatg gagactacaa
      241 agatcatgac attgattaca aggacgacga tgacaaataa gctagcttga ctgactgaga
      301 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      361 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      421 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      481 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      541 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
      601 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
      661 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
      721 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
      781 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
      841 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
      901 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
      961 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1021 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1921 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     1981 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2041 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2101 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2161 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2221 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2281 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     2341 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     2401 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     2461 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2521 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     2581 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2641 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2701 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2761 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2821 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     2881 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2941 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3001 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3061 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3121 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3181 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3241 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3301 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     3361 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     3421 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     3481 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     3541 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     3601 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     3661 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     3721 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     3781 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     3841 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3901 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3961 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4021 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4081 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4141 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4201 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4261 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4321 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4381 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     4441 gacacacccg ccagcggccg c
//
