LOCUS       sequence-294995-        4506 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 4506)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4506)
  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-m4134438
FEATURES             Location/Qualifiers
     source          1..4506
                     /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..322
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    375..496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(386..405)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     440..459
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      611..657
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      687..823
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    868..939
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1165..1753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1245..1264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2499..2567,2568..3359)
                     /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(2717..2736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3372..3508
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3660..3731
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3736..3782)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3800..3848
                     /note="synthetic polyadenylation signal"
     enhancer        3929..4232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4234..4436
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4387..4407
                     /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 tcagtcgaaa gaaaccgcgg cagctaagtt
      301 cgagaggcag cacatggaca gttaagctag cttgactgac tgagatacag cgtaccttca
      361 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      421 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      481 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      541 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
      601 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
      661 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
      721 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
      781 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
      841 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
      901 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
      961 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1021 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1081 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1981 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2041 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2101 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2161 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2221 gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     2281 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     2341 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     2401 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     2461 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     2521 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2581 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     2641 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     2701 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2761 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     2821 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2881 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     2941 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3001 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     3061 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     3121 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3181 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3241 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3301 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3361 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     3421 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     3481 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc
     3541 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg
     3601 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac
     3661 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc
     3721 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     3781 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
     3841 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat
     3901 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     3961 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4021 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4081 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4141 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4201 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg
     4261 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4321 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     4381 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     4441 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc
     4501 ggccgc
//
