LOCUS       sequence-300420-        6669 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 6669)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6669)
  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-m4252987
FEATURES             Location/Qualifiers
     source          1..6669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1183..1771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1672..1691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1997..2068
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2113..2249)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(2279..2325)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2440..2561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2477..2496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2531..2550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(2833..3035)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2862..2882)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3037..3340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3421..3469
                     /note="synthetic polyadenylation signal"
     terminator      3487..3533
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3538..3609
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3761..3897)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3910..4701,4702..4770))
                     /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     4533..4552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5036..5084
                     /note="synthetic polyadenylation signal"
     CDS             complement(5111..6139)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(join(6143..6669,1..683))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(6287..6306)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 aataagagct ttcccgcatt agcgaagggc tcaagacctt gggttcttgc cgcccaccat
       61 gccccccacc ttgtttcaac gacctcacag cccgcctcac aagcgtcttc cattcaagac
      121 tcgggaacag ccgccatttt gctgcgctcc ccccaacccc cagttcaggg caaccttgct
      181 cgcggaccca gactacagcc cttggcggtc tctccacacg cttccgtccc accgagcggc
      241 ccggcggcca cgaaagcccc ggccagccca gcagcccgct actcaccaag tgacgatcac
      301 agcgatccac aaacaagaac cgcgacccaa atcccggctg cgacggaact agctgtgcca
      361 cacccggcgc gtccttatat aatcatcggc gttcaccgcc ccacggagat ccctccgcag
      421 aatcgccgag aagggactac ttttcctcgc ctgttccgct ctctggaaag aaaaccagtg
      481 ccctagagtc acccaagtcc cgtcctaaaa tgtccttctg ctgatactgg ggttctaagg
      541 ccgagtctta tgagcagcgg gccgctgtcc tgagcgtccg ggcggaagga tcaggacgct
      601 cctgcgccct tcgtctgacg tggcagcgct cgccgtgacg aggggggcgc ccgcgggagg
      661 cgccaaaacc cggcgcggag gccaggcgcg ccccgatttc gatattcctc ggatctcgga
      721 tagatcggga ttgaggatgt aggttgaact tctctcgggt gatcccgatc tcggatattc
      781 gggattggag taatacctgg cgataggcgc tgccaagcaa tagccaaggc gcaaggccgg
      841 ccaagagact tgcgatagtt cgttcgagag gtagcctgca agatggatcg gacgaaaggt
      901 ttgaagattt tccgacgagg gccttggaga gttcggaaat ttaaatgcaa ctatggatga
      961 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1021 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1081 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1141 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1201 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1261 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1321 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1381 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1441 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1501 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1561 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1621 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1681 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1741 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1801 cctggccttt tgctggcctt ttgctcacat gatttaaatg aaactcgtaa tcggagaacg
     1861 ggtgcaatag caaggaccag atcgaacttg actgacgcaa ttctgagcgt agtcgaaacc
     1921 cagacaagtc gatctcgcca agtagctgac ttgactggtt aattaagttt cgatagccca
     1981 aggtaaccaa tgagattagg gagggaagaa agtttgatga actacttctg acccctagtg
     2041 gtgtccagaa aagaccatta aaggaatgtc ttgatcgaaa tcgagagatc gcctgcagga
     2101 tagctgacga ccttaaggat aaatttctgg taaggaggac acgtatggaa gtgggcaagt
     2161 tggggaagcc gtatccgttg ctgaatctgg catatgtggg agtataagac gcgcagcgtc
     2221 gcatcaggca tttttttctg cgccaatgcc ggtagatagc aatccggccc gaggggcaca
     2281 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctacgata ccgatagaga
     2341 tgggccaata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2401 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2521 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tgagctgaag
     2581 gtacgctgta tctcagtcag tcaagctagc agaggcctgc ggatccttag gcgtgacatg
     2641 tctcattatg atcttgtaac tccagactga accaaaaaat gattagcata atgagcacca
     2701 gaaaaagcag gaaagccaag aagcacagat aaaagtcgat gagcgatagt tctatcatgg
     2761 tggcggccgc tggcgggtgt gtcgagtgga tggtaggatc gacaaagatc tgatctgacg
     2821 gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     2881 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     2941 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     3001 tccacgccca ttgatgtact gccaaaaccg catcagcatg gtaatagcga tgactaatac
     3061 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     3121 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     3181 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     3241 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     3301 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     3361 atgaactaat gaccccgtaa ttgattacta gatctggcta cactcgatca gcagttagat
     3421 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgcaac
     3481 gattgatagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgaattgag
     3541 aggtagctga agctgctgtt atgaccacta gagggaagaa gatacctgtg gagctaatgg
     3601 tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa gctgacgata gctgacgaga
     3661 ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc gtaagctcga aacgacggag
     3721 attcgataga gccagagtcg agtttaaacg attcgggatc ttaaggataa atttctggta
     3781 aggaggacac gtatggaagt gggcaagttg gggaagccgt atccgttgct gaatctggca
     3841 tatgtgggag tataagacgc gcagcgtcgc atcaggcatt tttttctgcg ccaatgcacc
     3901 tagaatcggt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4141 ctccatccag tctatcaact gttgccggga agctagagta agtagttcgc cagttaatag
     4201 tttacgcaag gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4321 caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc agaagtaagt tggccgcagt
     4381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4441 atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt gtatgcggcg
     4501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4561 aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4621 gttgagatcc agttcgatgt aacccactcg cgcacccaac tgatcttcag catcttttac
     4681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4741 aagggcgaca cggaaatgtt gaatactcat gcgaaagcct cctgctgatc ctgtctcttg
     4801 atagatgttt gcaaaagcta ggactcgatt atacgttgac cgatcagact atcaatatct
     4861 aggtttaaac agatcggtac gctgccgtga ccaggcttga gtacaatatc tgacgattgg
     4921 acttcctggg tcccaagctg gattccgact ggctgactgc caagcaacga tatggatcgt
     4981 cgagactcgc actgagagga tcgttctcgg tacaccaggc gcgcctagcg ttacacacac
     5041 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa gactaggctt
     5101 acatactagc ttattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
     5161 cgagtacttc tacacagcca tcggtccaga cggcggcgct tctgcgggcg atttgtgtac
     5221 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     5281 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     5341 tatacgcccg gagccggggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     5401 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     5461 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
     5521 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
     5581 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
     5641 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
     5701 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
     5761 cggcggcagc gatggcatcc attgcctccg cgaccggctg cagaacagcg ggcagttcgg
     5821 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc
     5881 tctcgctgaa ctccccaatg tcaagcactt ccggaatcgg gagcgcggcg gatgcaaagt
     5941 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
     6001 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
     6061 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
     6121 gttcaggctt tttacccatg ttgtctaaca aaaaagccaa aaacggccag aatttagcgg
     6181 acaatttaca agtctaacac tgaaaattac atattgaccc aaatgattac atttcaaaag
     6241 gtgcctaaaa aacttcacaa aacacactcg ccaaccccga gcgcatagtt caaaaccgga
     6301 gcttcagcta cttaagaaga taggtacata aaaccgacca aagaaactga cgcctcactt
     6361 atccctcccc tcaccagagg tccggcgcct gtcgattcag gagagcctac cctaggcccg
     6421 aaccctgcgt cctgcgacgg agaaaagcct accgcacacc taccggctgg tggccccacc
     6481 ctgcattata agccaacaga acgggtgacg tcacgacacg acgagggcgc gcgctcccaa
     6541 aggtacgggt gcactgccca acggcaccgc cataactgcc gcccccgcaa cagacgacaa
     6601 accgagttct ccagtcagtg acaaacttca cgtcagggtc cccagatggt gcccagccca
     6661 tctcacccg
//
