LOCUS       sequence-295049-        5016 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 5016)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5016)
  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-m4134021
FEATURES             Location/Qualifiers
     source          1..5016
                     /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"
     CDS             join(3009..3077,3078..3869)
                     /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(3227..3246)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3882..4018
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4170..4241
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4246..4292)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4310..4358
                     /note="synthetic polyadenylation signal"
     enhancer        4439..4742
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4744..4946
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4897..4917
                     /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 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2821 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2881 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     2941 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     3001 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3061 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3121 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3181 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3241 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3301 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3361 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3421 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3481 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3541 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3601 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3661 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3721 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3781 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3841 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3901 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     3961 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     4021 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     4081 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     4141 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     4201 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     4261 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     4321 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     4381 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4441 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4501 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4561 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4621 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4681 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4741 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4801 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4861 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4921 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     4981 tcctaccatc cactcgacac acccgccagc ggccgc
//
