LOCUS       sequence-301654-        7134 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 7134)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7134)
  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-m4252942
FEATURES             Location/Qualifiers
     source          1..7134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..31
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(135..152)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(501..521)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(779..981)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(808..828)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        983..1286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1367..1415
                     /note="synthetic polyadenylation signal"
     terminator      1433..1479
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1484..1555
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1707..1843)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1856..2647,2648..2716))
                     /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     2479..2498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2982..3030
                     /note="synthetic polyadenylation signal"
     CDS             complement(3057..4085)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(4089..5298)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4233..4252)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5798..6386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6287..6306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6612..6683
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6728..6864)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6894..6940)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(7055..7134,1..42)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7092..7111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtgagctgaa
       61 ggtacgctgt atctcagtca gtcaagctag cagaggcctg cggatcctta gccgcgtggc
      121 acgagcccgc tgctatggtg atggtgatgg tgcatgtgcc ccgagcctga acctgccagg
      181 ttagcgtcca agaactcttt tagctggccc ttagacaggg ctcccacttt tgtcgcggca
      241 acttccccat tcttaaagag caggagggta ggtattcccc tgatgccgta tttaggtgcc
      301 gtccctgggt tctgatctat attaagctta gcgacagtta attttccttg atactcgtcg
      361 gcgatctcgt ccagaatggg tgcgatcatc ttgcacgggc cacaccattc agcccagaaa
      421 tctaccagta tcgcaccatc ggccttcaac acgtctgtat caaaggagtc atcggttaga
      481 tgaatgattt tgtcactcat cgactgaaag taaaggtttt catcaatctc cattggttgc
      541 tcttcatcta attgagaata tttattctca gttagtgact tagataaatt tttaattatg
      601 aggtttatga tgtaatcaag attccaaatg gaaactttaa aagtcctcat aataattagt
      661 aatatctctg ctatagtaac ctgaaagtca acgagatgaa acatggtggc ggccgctggc
      721 gggtgtgtcg agtggatggt aggatcgaca aagatctgat ctgacggttc actaaaccag
      781 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
      841 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
      901 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
      961 tgtactgcca aaaccgcatc agcatggtaa tagcgatgac taatacgtag atgtactgcc
     1021 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     1081 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     1141 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     1201 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     1261 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     1321 ccgtaattga ttactagatc tggctacact cgatcagcag ttagataata aaatcgctat
     1381 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa
     1441 ccccttgggg cctctaaacg ggtcttgagg ggttttttga attgagaggt agctgaagct
     1501 gctgttatga ccactagagg gaagaagata cctgtggagc taatggtcca agatgtcggg
     1561 agccgcgatc gctgccaaaa aggtaagctg acgatagctg acgagattcg atgcccgacg
     1621 agcaaatgag cgccacggct gttcgcgtaa gctcgaaacg acggagattc gatagagcca
     1681 gagtcgagtt taaacgattc gggatcttaa ggataaattt ctggtaagga ggacacgtat
     1741 ggaagtgggc aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata
     1801 agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa tgcacctaga atcggttacc
     1861 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1921 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1981 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2041 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2101 tcaactgttg ccgggaagct agagtaagta gttcgccagt taatagttta cgcaaggttg
     2161 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2221 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2281 gctccttcgg tcctccaatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2341 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2401 ctggtgaata ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2461 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2521 ttggaaagcg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2581 cgatgtaacc cactcgcgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2641 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2701 aatgttgaat actcatgcga aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa
     2761 aagctaggac tcgattatac gttgaccgat cagactatca atatctaggt ttaaacagat
     2821 cggtacgctg ccgtgaccag gcttgagtac aatatctgac gattggactt cctgggtccc
     2881 aagctggatt ccgactggct gactgccaag caacgatatg gatcgtcgag actcgcactg
     2941 agaggatcgt tctcggtaca ccaggcgcgc ctagcgttac acacacaaaa aaccaacaca
     3001 catccatctt cgatggatag cgattttatt atctaagact aggcttacat actagcttat
     3061 tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag tacttctaca
     3121 cagccatcgg tccagacggc ggcgcttctg cgggcgattt gtgtacgccc gacagtcccg
     3181 gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg
     3241 ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata cgcccggagc
     3301 cggggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg ctgctccata
     3361 caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga atccccgaac
     3421 atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag gacattgttg
     3481 gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc ccaaagcatc
     3541 agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac agtttgccag
     3601 tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt gtattgaccg
     3661 attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc ggcagcgatg
     3721 gcatccattg cctccgcgac cggctgcaga acagcgggca gttcggtttc aggcaggtct
     3781 tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc gctgaactcc
     3841 ccaatgtcaa gcacttccgg aatcgggagc gcggcggatg caaagtgccg ataaacataa
     3901 cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc acgccctcct
     3961 acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag gtcggagacg
     4021 ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc aggcttttta
     4081 cccatgttgt ctaacaaaaa agccaaaaac ggccagaatt tagcggacaa tttacaagtc
     4141 taacactgaa aattacatat tgacccaaat gattacattt caaaaggtgc ctaaaaaact
     4201 tcacaaaaca cactcgccaa ccccgagcgc atagttcaaa accggagctt cagctactta
     4261 agaagatagg tacataaaac cgaccaaaga aactgacgcc tcacttatcc ctcccctcac
     4321 cagaggtccg gcgcctgtcg attcaggaga gcctacccta ggcccgaacc ctgcgtcctg
     4381 cgacggagaa aagcctaccg cacacctacc ggctggtggc cccaccctgc attataagcc
     4441 aacagaacgg gtgacgtcac gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac
     4501 tgcccaacgg caccgccata actgccgccc ccgcaacaga cgacaaaccg agttctccag
     4561 tcagtgacaa acttcacgtc agggtcccca gatggtgccc agcccatctc acccgaataa
     4621 gagctttccc gcattagcga agggctcaag accttgggtt cttgccgccc accatgcccc
     4681 ccaccttgtt tcaacgacct cacagcccgc ctcacaagcg tcttccattc aagactcggg
     4741 aacagccgcc attttgctgc gctcccccca acccccagtt cagggcaacc ttgctcgcgg
     4801 acccagacta cagcccttgg cggtctctcc acacgcttcc gtcccaccga gcggcccggc
     4861 ggccacgaaa gccccggcca gcccagcagc ccgctactca ccaagtgacg atcacagcga
     4921 tccacaaaca agaaccgcga cccaaatccc ggctgcgacg gaactagctg tgccacaccc
     4981 ggcgcgtcct tatataatca tcggcgttca ccgccccacg gagatccctc cgcagaatcg
     5041 ccgagaaggg actacttttc ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta
     5101 gagtcaccca agtcccgtcc taaaatgtcc ttctgctgat actggggttc taaggccgag
     5161 tcttatgagc agcgggccgc tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc
     5221 gcccttcgtc tgacgtggca gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca
     5281 aaacccggcg cggaggccag gcgcgccccg atttcgatat tcctcggatc tcggatagat
     5341 cgggattgag gatgtaggtt gaacttctct cgggtgatcc cgatctcgga tattcgggat
     5401 tggagtaata cctggcgata ggcgctgcca agcaatagcc aaggcgcaag gccggccaag
     5461 agacttgcga tagttcgttc gagaggtagc ctgcaagatg gatcggacga aaggtttgaa
     5521 gattttccga cgagggcctt ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa
     5581 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5641 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5701 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5941 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6421 ccttttgctg gccttttgct cacatgattt aaatgaaact cgtaatcgga gaacgggtgc
     6481 aatagcaagg accagatcga acttgactga cgcaattctg agcgtagtcg aaacccagac
     6541 aagtcgatct cgccaagtag ctgacttgac tggttaatta agtttcgata gcccaaggta
     6601 accaatgaga ttagggaggg aagaaagttt gatgaactac ttctgacccc tagtggtgtc
     6661 cagaaaagac cattaaagga atgtcttgat cgaaatcgag agatcgcctg caggatagct
     6721 gacgacctta aggataaatt tctggtaagg aggacacgta tggaagtggg caagttgggg
     6781 aagccgtatc cgttgctgaa tctggcatat gtgggagtat aagacgcgca gcgtcgcatc
     6841 aggcattttt ttctgcgcca atgccggtag atagcaatcc ggcccgaggg gcacaaaaaa
     6901 cccctcaaga cccgtttaga ggccccaagg ggttatgcta cgataccgat agagatgggc
     6961 caataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     7021 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     7081 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cact
//
