LOCUS       sequence-299528-        6923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6923)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6923)
  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-m4258310
FEATURES             Location/Qualifiers
     source          1..6923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             311..388
                     /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             416..436
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    885..1006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(896..915)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     950..969
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1121..1167
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1197..1333
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1378..1449
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1675..2263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1755..1774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2763..3972
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3809..3828
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3976..4575
                     /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(3976..3995)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4472..4492
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4602..4650
                     /note="synthetic polyadenylation signal"
     CDS             join(4916..4984,4985..5776)
                     /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(5134..5153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5789..5925
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6077..6148
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6153..6199)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6217..6265
                     /note="synthetic polyadenylation signal"
     enhancer        6346..6649
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6651..6853
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6804..6824
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaagaaa acctttactt tcagtcggga cggcgaatcc cggggctgat
      301 aaatccctgg aaaaggagat ggaagaagaa ctttattgca gtgtctgcgg ctaaccggtt
      361 caaaaagatc agttcaagcg gcgccctaga ttacgacatt cctacgaccg cctccgagaa
      421 tctctatttc cagggtgaat ttggcttggc acaacatgac gaggccgtcg ataacaagtt
      481 taataaagaa cagcagaacg ctttctatga gatccttcac ttaccaaatc tgaacgaaga
      541 gcaacgcaac gctttcattc agagcctgaa agacgatccc tcgcagtctg caaatctcct
      601 ggccgaagcc aagaagctga atgacgcaca ggctccaaaa gttgataaca agtttaacaa
      661 ggagcagcag aatgctttct acgagatatt gcaccttcct aatctgaatg aggaacaaag
      721 aaatgcattt atccagtccc taaaagacga tccatcacag agtgccaacc tgcttgcgga
      781 agccaaaaag ctcaatgatg cccaggcccc gaaagtagac gcgaaccatc aataagctag
      841 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      901 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      961 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1021 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1081 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1141 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     1201 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1261 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1321 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1381 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1441 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1501 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1561 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2101 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2341 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2401 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2461 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     2521 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2581 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2641 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2701 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2761 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2821 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2881 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2941 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     3001 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     3061 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     3121 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     3181 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     3241 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     3301 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     3361 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     3421 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     3481 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     3541 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     3601 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg
     3661 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3721 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3781 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3841 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3901 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3961 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4021 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4081 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4141 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4201 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4261 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     4321 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4381 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     4441 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4501 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4561 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     4621 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4681 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4741 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4801 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4861 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4921 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4981 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     5041 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5101 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5161 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5221 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5281 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     5341 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5401 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5461 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     5521 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5581 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5641 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5701 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5761 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5821 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5881 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     5941 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     6001 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     6061 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     6121 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     6181 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     6241 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     6301 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6361 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6421 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6481 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6541 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6601 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6661 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6721 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6781 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6841 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     6901 tcgacacacc cgccagcggc cgc
//
