LOCUS       sequence-295023-        5100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5100)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5100)
  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-m4133385
FEATURES             Location/Qualifiers
     source          1..5100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             341..361
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             395..472
                     /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             500..520
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    969..1090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(980..999)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1034..1053
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1205..1251
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1281..1417
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1462..1533
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1759..2347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1839..1858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3093..3161,3162..3953)
                     /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(3311..3330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3966..4102
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4254..4325
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4330..4376)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4394..4442
                     /note="synthetic polyadenylation signal"
     enhancer        4523..4826
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4828..5030
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4981..5001
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat gagcttagtc ctgttgcact acgacagatg tcttgtgctg ccggtactac
       61 acaaactgct tgcactgatg acaatgcgtt agcttactac aacacaacaa agggaggtag
      121 gtttgtactt gcactgttat ccgatttaca ggatttgaaa tgggctagat tccctaagag
      181 tgatggaact ggtactatct atacagaact ggaaccacct tgtaggtttg ttacagacac
      241 acctaaaggt cctaaagtga agtatttata ctttattaaa ggattaaaca acctaaatag
      301 aggtatggta cttggtagtt tagctgccac agtacgtcta gaaaaccttt actttcagtc
      361 gggacggcga atcccggggc tgataaatcc ctggaaaagg agatggaaga agaactttat
      421 tgcagtgtct gcggctaacc ggttcaaaaa gatcagttca agcggcgccc tagattacga
      481 cattcctacg accgcctccg agaatctcta tttccagggt gaatttggct tggcacaaca
      541 tgacgaggcc gtcgataaca agtttaataa agaacagcag aacgctttct atgagatcct
      601 tcacttacca aatctgaacg aagagcaacg caacgctttc attcagagcc tgaaagacga
      661 tccctcgcag tctgcaaatc tcctggccga agccaagaag ctgaatgacg cacaggctcc
      721 aaaagttgat aacaagttta acaaggagca gcagaatgct ttctacgaga tattgcacct
      781 tcctaatctg aatgaggaac aaagaaatgc atttatccag tccctaaaag acgatccatc
      841 acagagtgcc aacctgcttg cggaagccaa aaagctcaat gatgcccagg ccccgaaagt
      901 agacgcgaac catcaataag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      961 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1021 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1081 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1141 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1201 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1261 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1321 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1381 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1441 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1501 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     1561 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     1621 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1681 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2461 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2521 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2581 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     2641 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2701 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2761 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2821 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     2881 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     2941 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     3001 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     3061 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     3121 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3181 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     3241 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     3301 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3361 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     3421 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3481 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     3541 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3601 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     3661 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     3721 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3781 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3841 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3901 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     3961 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     4021 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     4081 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     4141 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     4201 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     4261 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     4321 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     4381 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     4441 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     4501 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4561 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4621 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4681 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4741 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4801 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     4861 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4921 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4981 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     5041 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
