LOCUS       sequence-300632-        4461 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 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 Dec 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"
     misc_feature    284..355
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(400..536)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(566..612)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    727..848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(764..783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     818..837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(901..966)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(970..990)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(1248..1450)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1277..1297)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1452..1755
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1836..1884
                     /note="synthetic polyadenylation signal"
     terminator      1902..1948
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1953..2024
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2176..2312)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2325..3116,3117..3185))
                     /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     2948..2967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3931..4461,1..58)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4420..4439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgat
      121 ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa ggaccagatc gaacttgact
      181 gacgcaattc tgagcgtagt cgaaacccag acaagtcgat ctcgccaagt agctgacttg
      241 actggttaat taagtttcga tagcccaagg taaccaatga gattagggag ggaagaaagt
      301 ttgatgaact acttctgacc cctagtggtg tccagaaaag accattaaag gaatgtcttg
      361 atcgaaatcg agagatcgcc tgcaggatag ctgacgacct taaggataaa tttctggtaa
      421 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
      481 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgccggt
      541 agatagcaat ccggcccgag gggcacaaaa aacccctcaa gacccgttta gaggccccaa
      601 ggggttatgc tacgataccg atagagatgg gccaatacca catttgtaga ggttttactt
      661 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
      721 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
      781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
      841 gtatcttatc atgtctgtga gctgaaggta cgctgtatct cagtcagtca agctagctta
      901 tttgtcatcg tcgtccttgt aatcaatgtc atgatctttg tagtctccat cgtggtcctt
      961 ataatccatc gactgaaagt aaaggttttc atcaatctcc attggttgct cttcatctaa
     1021 ttgagaatat ttattctcag ttagtgactt agataaattt ttaattatga ggtttatgat
     1081 gtaatcaaga ttccaaatgg aaactttaaa agtcctcata ataattagta atatctctgc
     1141 tatagtaacc tgaaagtcaa cgagatgaaa catggtggcg gccgctggcg ggtgtgtcga
     1201 gtggatggta ggatcgacaa agatctgatc tgacggttca ctaaaccagc tctgcttata
     1261 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     1321 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     1381 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     1441 aaccgcatca gcatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     1501 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     1561 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     1621 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     1681 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     1741 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     1801 tactagatct ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat
     1861 ggatgtgtgt tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc
     1921 ctctaaacgg gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac
     1981 cactagaggg aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg
     2041 ctgccaaaaa ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc
     2101 gccacggctg ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt
     2161 aaacgattcg ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca
     2221 agttggggaa gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc
     2281 gtcgcatcag gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc
     2341 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2401 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2461 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2521 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc
     2581 cgggaagcta gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct
     2641 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2701 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2761 cctccaatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2821 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac
     2881 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2941 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt
     3001 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3061 actcgcgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3121 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3181 ctcatgcgaa agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact
     3241 cgattatacg ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc
     3301 cgtgaccagg cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc
     3361 cgactggctg actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt
     3421 ctcggtacac caggcgcgcc ccgatttcga tattcctcgg atctcggata gatcgggatt
     3481 gaggatgtag gttgaacttc tctcgggtga tcccgatctc ggatattcgg gattggagta
     3541 atacctggcg ataggcgctg ccaagcaata gccaaggcgc aaggccggcc aagagacttg
     3601 cgatagttcg ttcgagaggt agcctgcaag atggatcgga cgaaaggttt gaagattttc
     3661 cgacgagggc cttggagagt tcggaaattt aaatgcaact atggatgaac gaaatagaca
     3721 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3781 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3841 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4441 tagtcctgtc gggtttcgcc a
//
