LOCUS       sequence-300712-        4410 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 4410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4410)
  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-m4133215
FEATURES             Location/Qualifiers
     source          1..4410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    159..230
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(235..281)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    299..347
                     /note="synthetic polyadenylation signal"
     enhancer        428..731
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        733..935
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     886..906
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1193..1213
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1214..1231
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1284..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1295..1314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1349..1368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1520..1566
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1596..1732
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1777..1848
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2074..2662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2154..2173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3408..3476,3477..4268)
                     /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(3626..3645)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      join(4281..4410,1..7)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
       61 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
      121 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
      181 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
      241 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
      301 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
      361 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
      421 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      481 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      541 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      601 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      661 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      721 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      781 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      841 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      901 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
      961 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgccacca tgtttcatct
     1021 cgttgacttt caggttacta tagcagagat attactaatt attatgagga cttttaaagt
     1081 ttccatttgg aatcttgatt acatcataaa cctcataatt aaaaatttat ctaagtcact
     1141 aactgagaat aaatattctc aattagatga agagcaacca atggagattg atgaaaacct
     1201 ttactttcag tcgcaccatc accatcacca ttaagctagc ttgactgact gagatacagc
     1261 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1321 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1381 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1441 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1501 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1561 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1621 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1681 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1741 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1801 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1861 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1921 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1981 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     2041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2881 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2941 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     3001 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3061 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3121 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     3181 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     3241 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     3301 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     3361 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     3421 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3481 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     3541 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     3601 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3661 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     3721 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3781 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     3841 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3901 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3961 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     4021 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4081 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4141 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4201 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4261 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     4321 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     4381 tccatacgtg tcctccttac cagaaattta
//
