LOCUS       sequence-301656-        6290 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 6290)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6290)
  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-m4252779
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1458..2046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1947..1966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2272..2343
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2388..2524)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(2554..2600)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2715..2836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2752..2771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2806..2825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2889..2933)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(2934..2954)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3158..3360)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3187..3207)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3362..3665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3746..3794
                     /note="synthetic polyadenylation signal"
     terminator      3812..3858
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3863..3934
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4086..4222)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(4235..5026,5027..5095))
                     /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     4858..4877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5361..5409
                     /note="synthetic polyadenylation signal"
     CDS             complement(5436..6035)
                     /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(5519..5539)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6016..6035
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(6039..6290,1..958))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(6183..6202)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 attcaggaga gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg
       61 cacacctacc ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac
      121 gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata
      181 actgccgccc ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc
      241 agggtcccca gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga
      301 agggctcaag accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct
      361 cacagcccgc ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc
      421 gctcccccca acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg
      481 cggtctctcc acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca
      541 gcccagcagc ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga
      601 cccaaatccc ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca
      661 tcggcgttca ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc
      721 ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc
      781 taaaatgtcc ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc
      841 tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca
      901 gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag
      961 gcgcgccccg atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt
     1021 gaacttctct cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata
     1081 ggcgctgcca agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc
     1141 gagaggtagc ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt
     1201 ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa atagacagat cgctgagata
     1261 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1321 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1381 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1441 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1501 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1561 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1621 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1681 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1741 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1801 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1861 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1921 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1981 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2041 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2101 cacatgattt aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg accagatcga
     2161 acttgactga cgcaattctg agcgtagtcg aaacccagac aagtcgatct cgccaagtag
     2221 ctgacttgac tggttaatta agtttcgata gcccaaggta accaatgaga ttagggaggg
     2281 aagaaagttt gatgaactac ttctgacccc tagtggtgtc cagaaaagac cattaaagga
     2341 atgtcttgat cgaaatcgag agatcgcctg caggatagct gacgacctta aggataaatt
     2401 tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa
     2461 tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca
     2521 atgccggtag atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga
     2581 ggccccaagg ggttatgcta cgataccgat agagatgggc caataccaca tttgtagagg
     2641 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2701 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2821 tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag
     2881 ctagcttaac tgtccatgtg ctgcctctcg aacttagctg ccgcggtttc tttcgactga
     2941 aagtaaaggt tttcggcgtg acaagtttca ttatgatctt gcagttcaag tgagaaccaa
     3001 aagataataa gcataattaa aacaaggaat agcagaaagg ctaaaaagca caaatagaag
     3061 tcaattaatg aaagttcaat catggtggcg gccgctggcg ggtgtgtcga gtggatggta
     3121 ggatcgacaa agatctgatc tgacggttca ctaaaccagc tctgcttata tagacctccc
     3181 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     3241 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     3301 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     3361 gcatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     3421 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     3481 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     3541 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     3601 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     3661 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactagatct
     3721 ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat ggatgtgtgt
     3781 tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc ctctaaacgg
     3841 gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac cactagaggg
     3901 aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg ctgccaaaaa
     3961 ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc gccacggctg
     4021 ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt aaacgattcg
     4081 ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca agttggggaa
     4141 gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc gtcgcatcag
     4201 gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc agtgaggcac
     4261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc cgggaagcta
     4501 gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct acaggcatcg
     4561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccaatcg
     4681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac tcaaccaagt
     4801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt tcttcggggc
     4921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgcgcac
     4981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatgcgaa
     5101 agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact cgattatacg
     5161 ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc cgtgaccagg
     5221 cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc cgactggctg
     5281 actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt ctcggtacac
     5341 caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     5401 gattttatta tctaagacta ggcttacata ctagcctagg caccgggctt gcgggtcatg
     5461 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     5521 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
     5581 tcggcggcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     5641 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     5701 aggccctcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     5761 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     5821 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     5881 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     5941 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     6001 gtggcgaggc gcaccgtggg cttgtactcg gtcatgttgt ctaacaaaaa agccaaaaac
     6061 ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat tgacccaaat
     6121 gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa ccccgagcgc
     6181 atagttcaaa accggagctt cagctactta agaagatagg tacataaaac cgaccaaaga
     6241 aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg
//
