LOCUS       sequence-295430-        5106 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 5106)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5106)
  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-m4134306
FEATURES             Location/Qualifiers
     source          1..5106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             347..367
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             401..478
                     /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             506..526
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    975..1096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(986..1005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1040..1059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1211..1257
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1287..1423
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1468..1539
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1765..2353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1845..1864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3099..3167,3168..3959)
                     /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(3317..3336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3972..4108
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4260..4331
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4336..4382)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4400..4448
                     /note="synthetic polyadenylation signal"
     enhancer        4529..4832
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4834..5036
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4987..5007
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat aacgagttaa gcccagtagc gcttcgccag atgagttgcg ccgcaggtac
       61 aacccaaact gcttgtacgg atgacaacgc cctcgcttac tataatacca caaagggagg
      121 ccgattcgtt ctggccctgt tgtcggatct gcaagacttg aaatgggcac gttttcccaa
      181 gtccgacggg actggcacca tttacacaga actagagccg ccatgcaggt tcgtgactga
      241 tacccctaaa ggacccaagg tcaaatatct gtattttatc aaggggctta acaatctcaa
      301 tagaggcatg gtgttaggat ctctagccgc aacggtgcgg ctgcaagaaa acctttactt
      361 tcagtcggga cggcgaatcc cggggctgat aaatccctgg aaaaggagat ggaagaagaa
      421 ctttattgca gtgtctgcgg ctaaccggtt caaaaagatc agttcaagcg gcgccctaga
      481 ttacgacatt cctacgaccg cctccgagaa tctctatttc cagggtgaat ttggcttggc
      541 acaacatgac gaggccgtcg ataacaagtt taataaagaa cagcagaacg ctttctatga
      601 gatccttcac ttaccaaatc tgaacgaaga gcaacgcaac gctttcattc agagcctgaa
      661 agacgatccc tcgcagtctg caaatctcct ggccgaagcc aagaagctga atgacgcaca
      721 ggctccaaaa gttgataaca agtttaacaa ggagcagcag aatgctttct acgagatatt
      781 gcaccttcct aatctgaatg aggaacaaag aaatgcattt atccagtccc taaaagacga
      841 tccatcacag agtgccaacc tgcttgcgga agccaaaaag ctcaatgatg cccaggcccc
      901 gaaagtagac gcgaaccatc aataagctag cttgactgac tgagatacag cgtaccttca
      961 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1021 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1081 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1141 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     1201 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     1261 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     1321 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     1381 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     1441 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     1501 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     1561 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1621 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1681 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2581 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2641 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2701 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2761 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2821 gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     2881 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     2941 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     3001 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     3061 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     3121 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3181 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     3241 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     3301 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3361 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     3421 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3481 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     3541 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3601 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     3661 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     3721 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3781 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3841 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3901 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3961 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     4021 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     4081 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc
     4141 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg
     4201 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac
     4261 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc
     4321 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     4381 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
     4441 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat
     4501 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4561 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4621 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4681 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4741 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4801 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg
     4861 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4921 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     4981 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     5041 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc
     5101 ggccgc
//
