LOCUS       sequence-297454-        6717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6717)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6717)
  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-m4252947
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             155..181
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    250..371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(261..280)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     315..334
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      486..532
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      562..698
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    743..814
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1040..1628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1120..1139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2128..3337
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3174..3193
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3341..4369
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4396..4444
                     /note="synthetic polyadenylation signal"
     CDS             join(4710..4778,4779..5570)
                     /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(4928..4947)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5583..5719
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5871..5942
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5947..5993)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6011..6059
                     /note="synthetic polyadenylation signal"
     enhancer        6140..6443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6445..6647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6598..6618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgata gaactatcgc tcatcgactt ttatctgtgc ttcttggctt tcctgctttt
       61 tctggtgctc attatgctaa tcattttttg gttcagtctg gagttacaag atcataatga
      121 gacatgtcac gccgaaaacc tttactttca gtcgtacccc tatgacgtac cagattatgc
      181 gtaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
      241 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
      301 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
      361 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
      421 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
      481 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
      541 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
      601 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
      661 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
      721 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
      781 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
      841 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
      901 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
      961 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1021 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1081 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1141 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1201 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1261 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1321 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1381 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1441 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1501 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1561 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1621 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1681 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1741 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1801 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1861 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     1921 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     1981 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     2041 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     2101 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     2161 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     2221 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     2281 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     2341 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     2401 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     2461 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     2521 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     2581 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     2641 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     2701 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     2761 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     2821 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     2881 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     2941 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     3001 gtggggccac cagccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     3061 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     3121 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     3181 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     3241 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     3301 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     3361 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     3421 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     3481 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     3541 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     3601 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     3661 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     3721 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     3781 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     3841 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     3901 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     3961 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     4021 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     4081 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     4141 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     4201 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     4261 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     4321 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     4381 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     4441 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     4501 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     4561 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     4621 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     4681 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     4741 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4801 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     4861 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     4921 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4981 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     5041 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5101 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     5161 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5221 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     5281 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     5341 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5401 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5461 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5521 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     5581 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     5641 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     5701 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     5761 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     5821 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     5881 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     5941 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     6001 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     6061 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     6121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     6181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     6241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6421 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     6481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     6601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     6661 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
