LOCUS       sequence-295471-        4623 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 4623)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4623)
  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-m4133272
FEATURES             Location/Qualifiers
     source          1..4623
                     /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             362..439
                     /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    492..613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(503..522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     557..576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      728..774
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      804..940
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    985..1056
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1282..1870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1362..1381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2616..2684,2685..3476)
                     /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(2834..2853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3489..3625
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3777..3848
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3853..3899)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3917..3965
                     /note="synthetic polyadenylation signal"
     enhancer        4046..4349
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4351..4553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4504..4524
                     /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 gaagaggcga tggaaaaaga attttatcgc ggtgagtgca gccaacagat tcaaaaagat
      421 atcctcgtct ggtgccctct aagctagctt gactgactga gatacagcgt accttcagct
      481 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      541 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      601 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      661 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      721 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      781 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      841 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      901 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      961 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     1021 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     1081 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     1141 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     1201 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1681 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2101 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     2161 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     2221 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     2281 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     2341 ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     2401 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     2461 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     2521 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     2581 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     2641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2701 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     2761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     2821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2881 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     2941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3001 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     3061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3121 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     3181 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     3241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     3481 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     3541 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     3601 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     3661 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     3721 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     3781 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     3841 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     3901 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     3961 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     4021 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4081 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4141 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4201 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4261 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4321 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     4381 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4441 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4501 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     4561 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     4621 cgc
//
