LOCUS       sequence-300762-        7299 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 7299)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7299)
  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-m4252955
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        527..1736
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1573..1592
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1740..2768
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    2795..2843
                     /note="synthetic polyadenylation signal"
     CDS             join(3109..3177,3178..3969)
                     /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(3327..3346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3982..4118
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4270..4341
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4346..4392)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4410..4458
                     /note="synthetic polyadenylation signal"
     enhancer        4539..4842
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4844..5046
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4997..5017
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5304..5324
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5358..5435
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             5463..5483
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    5948..6069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5959..5978)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6013..6032
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      6184..6230
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      6260..6396
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6441..6512
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(join(6738..7299,1..27))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6818..6837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
       61 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      241 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
      301 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
      361 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
      421 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
      481 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg
      541 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa
      601 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
      661 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
      721 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
      781 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
      841 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
      901 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
      961 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     1021 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     1081 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     1141 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa
     1201 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca
     1261 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg
     1321 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc
     1381 tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc
     1441 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc
     1501 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc
     1561 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt
     1621 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt
     1681 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca
     1741 tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     1801 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     1861 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     1921 atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca
     1981 ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     2041 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg
     2101 atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     2161 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     2221 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     2281 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     2341 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     2401 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     2461 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     2521 cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     2581 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     2641 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg
     2701 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     2761 aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg
     2821 gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc
     2881 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     2941 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     3001 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     3061 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     3121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     3241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     3301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     3421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     3541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3661 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     3721 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3961 cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     4021 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     4081 ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg
     4141 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     4201 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     4261 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     4321 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     4381 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     4441 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     4501 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4561 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4621 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4681 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4741 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4801 acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag
     4861 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     4921 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     4981 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     5041 agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac
     5101 acccgccagc ggccgccacc atgttccact tggtggattt tcaggtcacg atcgcagaaa
     5161 tactgctaat tatcatgcga accttcaagg ttagcatttg gaacctggac tacatcatta
     5221 atctcatcat aaaaaacctt agtaagtctt taactgagaa taaatattcc caactggatg
     5281 aggaacagcc aatggagatt gacgaaaacc tttactttca gtcgggacgg cgaatcccgg
     5341 ggctgataaa tccctggaaa aggagatgga agaagaactt tattgcagtg tctgcggcta
     5401 accggttcaa aaagatcagt tcaagcggcg ccctagatta cgacattcct acgaccgcct
     5461 ccgagaatct ctatttccag ggtgaatttg gcttggcaca acatgacgag gccgtcgata
     5521 acaagtttaa taaagaacag cagaacgctt tctatgagat ccttcactta ccaaatctga
     5581 acgaagagca acgcaacgct ttcattcaga gcctgaaaga cgatccctcg cagtctgcaa
     5641 atctcctggc cgaagccaag aagctgaatg acgcacaggc tccaaaagtt gataacaagt
     5701 ttaacaagga gcagcagaat gctttctacg agatattgca ccttcctaat ctgaatgagg
     5761 aacaaagaaa tgcatttatc cagtccctaa aagacgatcc atcacagagt gccaacctgc
     5821 ttgcggaagc caaaaagctc aatgatgccc aggccccgaa agtagacgcg aaccatcaat
     5881 aaggatccgc aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca
     5941 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     6001 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     6061 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     6121 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     6181 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     6241 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     6301 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     6361 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga
     6421 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     6481 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa
     6541 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     6601 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     6661 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7141 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7201 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7261 accgctggta gcggtggttt ttttgtttgc aagcagcag
//
