LOCUS       sequence-300394-        6917 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 6917)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6917)
  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-m4252841
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             305..382
                     /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             410..430
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    879..1000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(890..909)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     944..963
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1115..1161
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1191..1327
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1372..1443
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1669..2257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1749..1768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2757..3966
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3803..3822
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3970..4569
                     /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(3970..3989)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4466..4486
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4596..4644
                     /note="synthetic polyadenylation signal"
     CDS             join(4910..4978,4979..5770)
                     /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(5128..5147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5783..5919
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6071..6142
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6147..6193)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6211..6259
                     /note="synthetic polyadenylation signal"
     enhancer        6340..6643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6645..6847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6798..6818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc gggacggcga atcccggggc tgataaatcc
      301 ctggaaaagg agatggaaga agaactttat tgcagtgtct gcggctaacc ggttcaaaaa
      361 gatcagttca agcggcgccc tagattacga cattcctacg accgcctccg agaatctcta
      421 tttccagggt gaatttggct tggcacaaca tgacgaggcc gtcgataaca agtttaataa
      481 agaacagcag aacgctttct atgagatcct tcacttacca aatctgaacg aagagcaacg
      541 caacgctttc attcagagcc tgaaagacga tccctcgcag tctgcaaatc tcctggccga
      601 agccaagaag ctgaatgacg cacaggctcc aaaagttgat aacaagttta acaaggagca
      661 gcagaatgct ttctacgaga tattgcacct tcctaatctg aatgaggaac aaagaaatgc
      721 atttatccag tccctaaaag acgatccatc acagagtgcc aacctgcttg cggaagccaa
      781 aaagctcaat gatgcccagg ccccgaaagt agacgcgaac catcaataag ctagcttgac
      841 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      901 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      961 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1021 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1081 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     1141 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     1201 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     1261 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     1321 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     1381 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1441 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1501 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1561 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2101 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2341 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2401 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2461 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2521 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2581 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2641 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2701 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2761 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2821 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2881 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2941 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     3001 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     3061 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     3121 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     3181 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     3241 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     3301 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     3361 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     3421 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     3481 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     3541 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3601 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     3661 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3721 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3781 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3841 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3901 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3961 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4021 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4081 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4141 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4201 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4261 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     4321 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4381 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     4441 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4501 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4561 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     4621 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4681 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4741 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4801 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4861 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4921 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4981 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     5041 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     5101 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5161 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     5221 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5281 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     5341 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5401 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5461 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5521 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5581 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5641 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5701 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5761 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5821 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5881 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5941 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     6001 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     6061 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     6121 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     6181 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     6241 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     6301 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6361 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6421 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6481 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6541 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6601 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     6661 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6721 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6781 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6841 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6901 cacccgccag cggccgc
//
