LOCUS       sequence-297459-        6446 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 6446)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6446)
  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-m4252848
FEATURES             Location/Qualifiers
     source          1..6446
                     /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             278..355
                     /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    408..529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(419..438)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     473..492
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      644..690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      720..856
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    901..972
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1198..1786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1278..1297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2286..3495
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3332..3351
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3499..4098
                     /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(3499..3518)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3995..4015
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4125..4173
                     /note="synthetic polyadenylation signal"
     CDS             join(4439..4507,4508..5299)
                     /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(4657..4676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5312..5448
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5600..5671
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5676..5722)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5740..5788
                     /note="synthetic polyadenylation signal"
     enhancer        5869..6172
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6174..6376
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6327..6347
                     /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 tcagtcgaag aggcgatgga aaaagaattt
      301 tatcgcggtg agtgcagcca acagattcaa aaagatatcc tcgtctggtg ccctctaagc
      361 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      421 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      481 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      541 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
      601 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
      661 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
      721 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
      781 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
      841 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
      901 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
      961 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1021 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1081 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2041 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2101 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2161 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2221 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2281 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     2341 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     2401 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     2461 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     2521 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     2581 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     2641 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     2701 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     2761 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     2821 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     2881 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     2941 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     3001 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     3061 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3121 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca gccggtaggt
     3181 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     3241 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     3301 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     3361 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     3421 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     3481 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     3541 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     3601 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     3661 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     3721 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     3781 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     3841 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     3901 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     3961 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4021 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4081 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     4141 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     4201 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     4261 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     4321 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     4381 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     4441 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4501 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     4561 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4621 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4681 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4741 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4801 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     4861 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4921 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4981 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     5041 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5101 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5161 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5221 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5281 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     5341 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5401 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     5461 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     5521 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     5581 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     5641 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     5701 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     5761 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     5821 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5881 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5941 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6001 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6061 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6121 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     6181 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     6241 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     6301 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6361 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
     6421 cactcgacac acccgccagc ggccgc
//
