LOCUS       sequence-307372-        6287 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 6287)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6287)
  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-m4252833
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..123)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      1669..2257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2158..2177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2483..2554
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2599..2735)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(2765..2811)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2926..3047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2963..2982)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3017..3036
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3100..3141)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3142..3162)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3366..3568)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3395..3415)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3570..3873
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3954..4002
                     /note="synthetic polyadenylation signal"
     terminator      4020..4066
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    4071..4142
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4294..4430)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(4443..5234,5235..5303))
                     /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     5066..5085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5569..5617
                     /note="synthetic polyadenylation signal"
     CDS             complement(5644..6243)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5727..5747)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6224..6243
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(6247..6287,1..1169))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 atttacaagt ctaacactga aaattacata ttgacccaaa tgattacatt tcaaaaggtg
       61 cctaaaaaac ttcacaaaac acactcgcca accccgagcg catagttcaa aaccggagct
      121 tcagctactt aagaagatag gtacataaaa ccgaccaaag aaactgacgc ctcacttatc
      181 cctcccctca ccagaggtcc ggcgcctgtc gattcaggag agcctaccct aggcccgaac
      241 cctgcgtcct gcgacggaga aaagcctacc gcacacctac cggctggtgg ccccaccctg
      301 cattataagc caacagaacg ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg
      361 tacgggtgca ctgcccaacg gcaccgccat aactgccgcc cccgcaacag acgacaaacc
      421 gagttctcca gtcagtgaca aacttcacgt cagggtcccc agatggtgcc cagcccatct
      481 cacccgaata agagctttcc cgcattagcg aagggctcaa gaccttgggt tcttgccgcc
      541 caccatgccc cccaccttgt ttcaacgacc tcacagcccg cctcacaagc gtcttccatt
      601 caagactcgg gaacagccgc cattttgctg cgctcccccc aacccccagt tcagggcaac
      661 cttgctcgcg gacccagact acagcccttg gcggtctctc cacacgcttc cgtcccaccg
      721 agcggcccgg cggccacgaa agccccggcc agcccagcag cccgctactc accaagtgac
      781 gatcacagcg atccacaaac aagaaccgcg acccaaatcc cggctgcgac ggaactagct
      841 gtgccacacc cggcgcgtcc ttatataatc atcggcgttc accgccccac ggagatccct
      901 ccgcagaatc gccgagaagg gactactttt cctcgcctgt tccgctctct ggaaagaaaa
      961 ccagtgccct agagtcaccc aagtcccgtc ctaaaatgtc cttctgctga tactggggtt
     1021 ctaaggccga gtcttatgag cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag
     1081 gacgctcctg cgcccttcgt ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc
     1141 gggaggcgcc aaaacccggc gcggaggcca ggcgcgcccc gatttcgata ttcctcggat
     1201 ctcggataga tcgggattga ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg
     1261 atattcggga ttggagtaat acctggcgat aggcgctgcc aagcaatagc caaggcgcaa
     1321 ggccggccaa gagacttgcg atagttcgtt cgagaggtag cctgcaagat ggatcggacg
     1381 aaaggtttga agattttccg acgagggcct tggagagttc ggaaatttaa atgcaactat
     1441 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1501 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2281 acggttcctg gccttttgct ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg
     2341 agaacgggtg caatagcaag gaccagatcg aacttgactg acgcaattct gagcgtagtc
     2401 gaaacccaga caagtcgatc tcgccaagta gctgacttga ctggttaatt aagtttcgat
     2461 agcccaaggt aaccaatgag attagggagg gaagaaagtt tgatgaacta cttctgaccc
     2521 ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga tcgaaatcga gagatcgcct
     2581 gcaggatagc tgacgacctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
     2641 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
     2701 agcgtcgcat caggcatttt tttctgcgcc aatgccggta gatagcaatc cggcccgagg
     2761 ggcacaaaaa acccctcaag acccgtttag aggccccaag gggttatgct acgataccga
     2821 tagagatggg ccaataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2881 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtgag
     3061 ctgaaggtac gctgtatctc agtcagtcaa gctagcttag gtagaatcta agccgagaag
     3121 tggattgggt ataggtttcc ccgactgaaa gtaaaggttt tcggcgtgac atgtctcatt
     3181 atgatcttgt aactccagac tgaaccaaaa aatgattagc ataatgagca ccagaaaaag
     3241 caggaaagcc aagaagcaca gataaaagtc gatgagcgat agttctatca tggtggcggc
     3301 cgctggcggg tgtgtcgagt ggatggtagg atcgacaaag atctgatctg acggttcact
     3361 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     3421 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     3481 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     3541 ccattgatgt actgccaaaa ccgcatcagc atggtaatag cgatgactaa tacgtagatg
     3601 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     3661 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     3721 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     3781 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3841 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3901 aatgaccccg taattgatta ctagatctgg ctacactcga tcagcagtta gataataaaa
     3961 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat
     4021 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgaatt gagaggtagc
     4081 tgaagctgct gttatgacca ctagagggaa gaagatacct gtggagctaa tggtccaaga
     4141 tgtcgggagc cgcgatcgct gccaaaaagg taagctgacg atagctgacg agattcgatg
     4201 cccgacgagc aaatgagcgc cacggctgtt cgcgtaagct cgaaacgacg gagattcgat
     4261 agagccagag tcgagtttaa acgattcggg atcttaagga taaatttctg gtaaggagga
     4321 cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg
     4381 gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc acctagaatc
     4441 ggttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4681 cagtctatca actgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttacgc
     4741 aaggttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4861 gcggttagct ccttcggtcc tccaatcgtt gtcagaagta agttggccgc agtgttatca
     4921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4981 tctgtgactg gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5101 ctcatcattg gaaagcgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5161 tccagttcga tgtaacccac tcgcgcaccc aactgatctt cagcatcttt tactttcacc
     5221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5281 acacggaaat gttgaatact catgcgaaag cctcctgctg atcctgtctc ttgatagatg
     5341 tttgcaaaag ctaggactcg attatacgtt gaccgatcag actatcaata tctaggttta
     5401 aacagatcgg tacgctgccg tgaccaggct tgagtacaat atctgacgat tggacttcct
     5461 gggtcccaag ctggattccg actggctgac tgccaagcaa cgatatggat cgtcgagact
     5521 cgcactgaga ggatcgttct cggtacacca ggcgcgccta gcgttacaca cacaaaaaac
     5581 caacacacat ccatcttcga tggatagcga ttttattatc taagactagg cttacatact
     5641 agcctaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac
     5701 gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt
     5761 ctccaggaag gcgggcaccc cggcgcgctc ggcggcctcc actccgggga gcacgacggc
     5821 gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc
     5881 gggctccttg ggccggtgcg gcgccaggag gccctccatc tgttgctgcg cggccagccg
     5941 ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac
     6001 gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc
     6061 gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg
     6121 gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt
     6181 gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt
     6241 catgttgtct aacaaaaaag ccaaaaacgg ccagaattta gcggaca
//
