LOCUS       sequence-297303-        6323 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 6323)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6323)
  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-m4252866
FEATURES             Location/Qualifiers
     source          1..6323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             155..232
                     /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"
     polyA_signal    285..406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(296..315)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     350..369
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      521..567
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      597..733
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    778..849
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1075..1663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1155..1174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2163..3372
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3209..3228
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3376..3975
                     /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(3376..3395)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3872..3892
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4002..4050
                     /note="synthetic polyadenylation signal"
     CDS             join(4316..4384,4385..5176)
                     /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     complement(4534..4553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5189..5325
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5477..5548
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5553..5599)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5617..5665
                     /note="synthetic polyadenylation signal"
     enhancer        5746..6049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6051..6253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6204..6224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgata gaactatcgc tcatcgactt ttatctgtgc ttcttggctt tcctgctttt
       61 tctggtgctc attatgctaa tcattttttg gttcagtctg gagttacaag atcataatga
      121 gacatgtcac gccgaaaacc tttactttca gtcgaagagg cgatggaaaa agaattttat
      181 cgcggtgagt gcagccaaca gattcaaaaa gatatcctcg tctggtgccc tctaagctag
      241 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      301 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      361 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      421 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      481 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      541 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      601 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
      661 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
      721 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
      781 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
      841 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
      901 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
      961 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1861 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     1921 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     1981 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2041 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2101 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2161 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2221 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2281 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2341 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     2401 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     2461 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     2521 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     2581 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     2641 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     2701 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     2761 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     2821 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     2881 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     2941 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     3001 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg
     3061 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3121 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3181 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3241 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3301 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3361 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3421 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3481 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3541 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3601 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3661 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     3721 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3781 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     3841 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3901 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     3961 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     4021 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4081 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4141 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4201 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4261 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     4441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4501 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4621 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     4741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4861 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     4921 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5161 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5221 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5281 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     5341 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     5401 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     5461 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     5521 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     5581 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     5641 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     5701 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5761 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5821 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5881 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5941 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6001 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6061 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6121 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6181 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6241 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     6301 tcgacacacc cgccagcggc cgc
//
