LOCUS       sequence-300567-        6800 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 6800)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6800)
  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-m4252842
FEATURES             Location/Qualifiers
     source          1..6800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    107..155
                     /note="synthetic polyadenylation signal"
     terminator      173..219
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    224..295
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(447..583)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(596..1387,1388..1456))
                     /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     1219..1238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1722..1770
                     /note="synthetic polyadenylation signal"
     CDS             complement(1797..2396)
                     /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(1880..1900)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2377..2396
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2400..3609)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2544..2563)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4109..4697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4598..4617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4923..4994
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5039..5175)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5205..5251)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5366..5487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5403..5422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5457..5476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5936..5956)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5984..6061)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             complement(6095..6115)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6319..6521)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6348..6368)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6523..6800,1..26)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
       61 ccgtaattga ttactagatc tggctacact cgatcagcag ttagataata aaatcgctat
      121 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa
      181 ccccttgggg cctctaaacg ggtcttgagg ggttttttga attgagaggt agctgaagct
      241 gctgttatga ccactagagg gaagaagata cctgtggagc taatggtcca agatgtcggg
      301 agccgcgatc gctgccaaaa aggtaagctg acgatagctg acgagattcg atgcccgacg
      361 agcaaatgag cgccacggct gttcgcgtaa gctcgaaacg acggagattc gatagagcca
      421 gagtcgagtt taaacgattc gggatcttaa ggataaattt ctggtaagga ggacacgtat
      481 ggaagtgggc aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata
      541 agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa tgcacctaga atcggttacc
      601 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      661 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      721 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      781 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      841 tcaactgttg ccgggaagct agagtaagta gttcgccagt taatagttta cgcaaggttg
      901 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      961 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1021 gctccttcgg tcctccaatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1081 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1141 ctggtgaata ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1201 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1261 ttggaaagcg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1321 cgatgtaacc cactcgcgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1381 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1441 aatgttgaat actcatgcga aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa
     1501 aagctaggac tcgattatac gttgaccgat cagactatca atatctaggt ttaaacagat
     1561 cggtacgctg ccgtgaccag gcttgagtac aatatctgac gattggactt cctgggtccc
     1621 aagctggatt ccgactggct gactgccaag caacgatatg gatcgtcgag actcgcactg
     1681 agaggatcgt tctcggtaca ccaggcgcgc ctagcgttac acacacaaaa aaccaacaca
     1741 catccatctt cgatggatag cgattttatt atctaagact aggcttacat actagcctag
     1801 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     1861 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     1921 aaggcgggca ccccggcgcg ctcggcggcc tccactccgg ggagcacgac ggcgctgccc
     1981 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     2041 ttgggccggt gcggcgccag gaggccctcc atctgttgct gcgcggccag ccgggaaccg
     2101 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     2161 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     2221 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     2281 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     2341 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatgttg
     2401 tctaacaaaa aagccaaaaa cggccagaat ttagcggaca atttacaagt ctaacactga
     2461 aaattacata ttgacccaaa tgattacatt tcaaaaggtg cctaaaaaac ttcacaaaac
     2521 acactcgcca accccgagcg catagttcaa aaccggagct tcagctactt aagaagatag
     2581 gtacataaaa ccgaccaaag aaactgacgc ctcacttatc cctcccctca ccagaggtcc
     2641 ggcgcctgtc gattcaggag agcctaccct aggcccgaac cctgcgtcct gcgacggaga
     2701 aaagcctacc gcacacctac cggctggtgg ccccaccctg cattataagc caacagaacg
     2761 ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg tacgggtgca ctgcccaacg
     2821 gcaccgccat aactgccgcc cccgcaacag acgacaaacc gagttctcca gtcagtgaca
     2881 aacttcacgt cagggtcccc agatggtgcc cagcccatct cacccgaata agagctttcc
     2941 cgcattagcg aagggctcaa gaccttgggt tcttgccgcc caccatgccc cccaccttgt
     3001 ttcaacgacc tcacagcccg cctcacaagc gtcttccatt caagactcgg gaacagccgc
     3061 cattttgctg cgctcccccc aacccccagt tcagggcaac cttgctcgcg gacccagact
     3121 acagcccttg gcggtctctc cacacgcttc cgtcccaccg agcggcccgg cggccacgaa
     3181 agccccggcc agcccagcag cccgctactc accaagtgac gatcacagcg atccacaaac
     3241 aagaaccgcg acccaaatcc cggctgcgac ggaactagct gtgccacacc cggcgcgtcc
     3301 ttatataatc atcggcgttc accgccccac ggagatccct ccgcagaatc gccgagaagg
     3361 gactactttt cctcgcctgt tccgctctct ggaaagaaaa ccagtgccct agagtcaccc
     3421 aagtcccgtc ctaaaatgtc cttctgctga tactggggtt ctaaggccga gtcttatgag
     3481 cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag gacgctcctg cgcccttcgt
     3541 ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc gggaggcgcc aaaacccggc
     3601 gcggaggcca ggcgcgcccc gatttcgata ttcctcggat ctcggataga tcgggattga
     3661 ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg atattcggga ttggagtaat
     3721 acctggcgat aggcgctgcc aagcaatagc caaggcgcaa ggccggccaa gagacttgcg
     3781 atagttcgtt cgagaggtag cctgcaagat ggatcggacg aaaggtttga agattttccg
     3841 acgagggcct tggagagttc ggaaatttaa atgcaactat ggatgaacga aatagacaga
     3901 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3961 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4021 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg agaacgggtg caatagcaag
     4801 gaccagatcg aacttgactg acgcaattct gagcgtagtc gaaacccaga caagtcgatc
     4861 tcgccaagta gctgacttga ctggttaatt aagtttcgat agcccaaggt aaccaatgag
     4921 attagggagg gaagaaagtt tgatgaacta cttctgaccc ctagtggtgt ccagaaaaga
     4981 ccattaaagg aatgtcttga tcgaaatcga gagatcgcct gcaggatagc tgacgacctt
     5041 aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg gaagccgtat
     5101 ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat caggcatttt
     5161 tttctgcgcc aatgccggta gatagcaatc cggcccgagg ggcacaaaaa acccctcaag
     5221 acccgtttag aggccccaag gggttatgct acgataccga tagagatggg ccaataccac
     5281 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5341 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5401 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5461 tttgtccaaa ctcatcaatg tatcttatca tgtctgtgag ctgaaggtac gctgtatctc
     5521 agtcagtcaa gctagcttat tgatggttcg cgtctacttt cggggcctgg gcatcattga
     5581 gctttttggc ttccgcaagc aggttggcac tctgtgatgg atcgtctttt agggactgga
     5641 taaatgcatt tctttgttcc tcattcagat taggaaggtg caatatctcg tagaaagcat
     5701 tctgctgctc cttgttaaac ttgttatcaa cttttggagc ctgtgcgtca ttcagcttct
     5761 tggcttcggc caggagattt gcagactgcg agggatcgtc tttcaggctc tgaatgaaag
     5821 cgttgcgttg ctcttcgttc agatttggta agtgaaggat ctcatagaaa gcgttctgct
     5881 gttctttatt aaacttgtta tcgacggcct cgtcatgttg tgccaagcca aattcaccct
     5941 ggaaatagag attctcggag gcggtcgtag gaatgtcgta atctagggcg ccgcttgaac
     6001 tgatcttttt gaaccggtta gccgcagaca ctgcaataaa gttcttcttc catctccttt
     6061 tccagggatt tatcagcccc gggattcgcc gtcccgactg aaagtaaagg ttttcggcgt
     6121 gacaagtttc attatgatct tgcagttcaa gtgagaacca aaagataata agcataatta
     6181 aaacaaggaa tagcagaaag gctaaaaagc acaaatagaa gtcaattaat gaaagttcaa
     6241 tcatggtggc ggccgctggc gggtgtgtcg agtggatggt aggatcgaca aagatctgat
     6301 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     6361 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     6421 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     6481 ccgctatcca cgcccattga tgtactgcca aaaccgcatc agcatggtaa tagcgatgac
     6541 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     6601 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     6661 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     6721 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     6781 gttgggcggt cagccaggcg
//
