LOCUS       sequence-300568-        6635 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 6635)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6635)
  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-m4252850
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    21..69
                     /note="synthetic polyadenylation signal"
     terminator      87..133
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    138..209
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(361..497)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(510..1301,1302..1370))
                     /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     1133..1152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1636..1684
                     /note="synthetic polyadenylation signal"
     CDS             complement(1711..2310)
                     /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(1794..1814)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2291..2310
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2314..3523)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2458..2477)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4023..4611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4512..4531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4837..4908
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4953..5089)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5119..5165)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5280..5401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5317..5336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5371..5390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5478..5495)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5844..5864)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6068..6270)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6097..6117)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6272..6575
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cactcgatca gcagttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
       61 ttttgtgtgt gtaacgcaac gattgatagc ataacccctt ggggcctcta aacgggtctt
      121 gaggggtttt ttgaattgag aggtagctga agctgctgtt atgaccacta gagggaagaa
      181 gatacctgtg gagctaatgg tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa
      241 gctgacgata gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc
      301 gtaagctcga aacgacggag attcgataga gccagagtcg agtttaaacg attcgggatc
      361 ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt
      421 atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt
      481 tttttctgcg ccaatgcacc tagaatcggt taccaatgct taatcagtga ggcacctatc
      541 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      601 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      661 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      721 ggtcctgcaa ctttatccgc ctccatccag tctatcaact gttgccggga agctagagta
      781 agtagttcgc cagttaatag tttacgcaag gttgttgcca ttgctacagg catcgtggtg
      841 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      901 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc
      961 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1021 actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc
     1081 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1141 gcgccacata gcagaacttt aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa
     1201 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg cgcacccaac
     1261 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1321 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat gcgaaagcct
     1381 cctgctgatc ctgtctcttg atagatgttt gcaaaagcta ggactcgatt atacgttgac
     1441 cgatcagact atcaatatct aggtttaaac agatcggtac gctgccgtga ccaggcttga
     1501 gtacaatatc tgacgattgg acttcctggg tcccaagctg gattccgact ggctgactgc
     1561 caagcaacga tatggatcgt cgagactcgc actgagagga tcgttctcgg tacaccaggc
     1621 gcgcctagcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     1681 tattatctaa gactaggctt acatactagc ctaggcaccg ggcttgcggg tcatgcacca
     1741 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     1801 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     1861 ggcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     1921 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     1981 ctccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     2041 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     2101 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     2161 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     2221 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     2281 gaggcgcacc gtgggcttgt actcggtcat gttgtctaac aaaaaagcca aaaacggcca
     2341 gaatttagcg gacaatttac aagtctaaca ctgaaaatta catattgacc caaatgatta
     2401 catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg agcgcatagt
     2461 tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc aaagaaactg
     2521 acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca ggagagccta
     2581 ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac ctaccggctg
     2641 gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac gacgagggcg
     2701 cgcgctccca aaggtacggg tgcactgccc aacggcaccg ccataactgc cgcccccgca
     2761 acagacgaca aaccgagttc tccagtcagt gacaaacttc acgtcagggt ccccagatgg
     2821 tgcccagccc atctcacccg aataagagct ttcccgcatt agcgaagggc tcaagacctt
     2881 gggttcttgc cgcccaccat gccccccacc ttgtttcaac gacctcacag cccgcctcac
     2941 aagcgtcttc cattcaagac tcgggaacag ccgccatttt gctgcgctcc ccccaacccc
     3001 cagttcaggg caaccttgct cgcggaccca gactacagcc cttggcggtc tctccacacg
     3061 cttccgtccc accgagcggc ccggcggcca cgaaagcccc ggccagccca gcagcccgct
     3121 actcaccaag tgacgatcac agcgatccac aaacaagaac cgcgacccaa atcccggctg
     3181 cgacggaact agctgtgcca cacccggcgc gtccttatat aatcatcggc gttcaccgcc
     3241 ccacggagat ccctccgcag aatcgccgag aagggactac ttttcctcgc ctgttccgct
     3301 ctctggaaag aaaaccagtg ccctagagtc acccaagtcc cgtcctaaaa tgtccttctg
     3361 ctgatactgg ggttctaagg ccgagtctta tgagcagcgg gccgctgtcc tgagcgtccg
     3421 ggcggaagga tcaggacgct cctgcgccct tcgtctgacg tggcagcgct cgccgtgacg
     3481 aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag gccaggcgcg ccccgatttc
     3541 gatattcctc ggatctcgga tagatcggga ttgaggatgt aggttgaact tctctcgggt
     3601 gatcccgatc tcggatattc gggattggag taatacctgg cgataggcgc tgccaagcaa
     3661 tagccaaggc gcaaggccgg ccaagagact tgcgatagtt cgttcgagag gtagcctgca
     3721 agatggatcg gacgaaaggt ttgaagattt tccgacgagg gccttggaga gttcggaaat
     3781 ttaaatgcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gatttaaatg
     4681 aaactcgtaa tcggagaacg ggtgcaatag caaggaccag atcgaacttg actgacgcaa
     4741 ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca agtagctgac ttgactggtt
     4801 aattaagttt cgatagccca aggtaaccaa tgagattagg gagggaagaa agtttgatga
     4861 actacttctg acccctagtg gtgtccagaa aagaccatta aaggaatgtc ttgatcgaaa
     4921 tcgagagatc gcctgcagga tagctgacga ccttaaggat aaatttctgg taaggaggac
     4981 acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg
     5041 agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgcc ggtagatagc
     5101 aatccggccc gaggggcaca aaaaacccct caagacccgt ttagaggccc caaggggtta
     5161 tgctacgata ccgatagaga tgggccaata ccacatttgt agaggtttta cttgctttaa
     5221 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5401 atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc ttagccgcgt
     5461 ggcacgagcc cgctgctatg gtgatggtga tggtgcatgt gccccgagcc tgaacctgcc
     5521 aggttagcgt ccaagaactc ttttagctgg cccttagaca gggctcccac ttttgtcgcg
     5581 gcaacttccc cattcttaaa gagcaggagg gtaggtattc ccctgatgcc gtatttaggt
     5641 gccgtccctg ggttctgatc tatattaagc ttagcgacag ttaattttcc ttgatactcg
     5701 tcggcgatct cgtccagaat gggtgcgatc atcttgcacg ggccacacca ttcagcccag
     5761 aaatctacca gtatcgcacc atcggccttc aacacgtctg tatcaaagga gtcatcggtt
     5821 agatgaatga ttttgtcact catcgactga aagtaaaggt tttcggcgtg acaagtttca
     5881 ttatgatctt gcagttcaag tgagaaccaa aagataataa gcataattaa aacaaggaat
     5941 agcagaaagg ctaaaaagca caaatagaag tcaattaatg aaagttcaat catggtggcg
     6001 gccgctggcg ggtgtgtcga gtggatggta ggatcgacaa agatctgatc tgacggttca
     6061 ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     6121 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     6181 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     6241 gcccattgat gtactgccaa aaccgcatca gcatggtaat agcgatgact aatacgtaga
     6301 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     6361 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     6421 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     6481 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     6541 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     6601 ctaatgaccc cgtaattgat tactagatct ggcta
//
