LOCUS       sequence-295460-        5637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5637)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5637)
  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-m4134163
FEATURES             Location/Qualifiers
     source          1..5637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             878..898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             932..1009
                     /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             1037..1057
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1506..1627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1517..1536)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1571..1590
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1742..1788
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1818..1954
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1999..2070
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2296..2884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2376..2395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3630..3698,3699..4490)
                     /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(3848..3867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4503..4639
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4791..4862
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4867..4913)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4931..4979
                     /note="synthetic polyadenylation signal"
     enhancer        5060..5363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5365..5567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5518..5538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc gccgttaagc ggacaatcaa agggacccat cactggttgc tccttactat
       61 actgacgtct ttactggtct tggtacagag tacacaatgg tcgcttttct tttttctgta
      121 cgagaatgca ttcctcccat ttgcgatggg aatcattgct atgtccgcct tcgctatgat
      181 gttcgtgaag cacaaacatg ccttcctttg cctgtttctc ctaccctcac tggcaaccgt
      241 ggcctatttt aacatggtgt acatgcctgc gagctgggtc atgcgaatta tgacttggct
      301 ggacatggtt gatacctccc tatctggctt caagttaaag gactgtgtga tgtatgcatc
      361 agctgtagtg ctgctaatcc tgatgacagc tcgcacagtc tacgatgacg gtgccaggag
      421 agtgtggact ctgatgaatg tcctcacctt agtttataaa gtatactatg gcaacgctct
      481 cgatcaggcc attagtatgt gggcattgat aatctccgtg acgagtaact acagcggagt
      541 ggtaaccact gtcatgtttt tggcacgcgg gatagtgttc atgtgtgtcg aatattgccc
      601 gatctttttt atcacaggga atactctgca atgcattatg ctggtttact gtttcttggg
      661 ctatttctgc acctgttatt ttggtctatt ctgcctgctt aataggtact ttcgtttgac
      721 actaggagtc tatgattact tggtgagcac gcaggagttt cggtatatga acagtcaggg
      781 cctgcttccc ccaaagaact ctattgacgc gtttaaactc aatataaagc tgttgggcgt
      841 gggtggaaaa ccttgcatca aggtggccac tgtgcaggaa aacctttact ttcagtcggg
      901 acggcgaatc ccggggctga taaatccctg gaaaaggaga tggaagaaga actttattgc
      961 agtgtctgcg gctaaccggt tcaaaaagat cagttcaagc ggcgccctag attacgacat
     1021 tcctacgacc gcctccgaga atctctattt ccagggtgaa tttggcttgg cacaacatga
     1081 cgaggccgtc gataacaagt ttaataaaga acagcagaac gctttctatg agatccttca
     1141 cttaccaaat ctgaacgaag agcaacgcaa cgctttcatt cagagcctga aagacgatcc
     1201 ctcgcagtct gcaaatctcc tggccgaagc caagaagctg aatgacgcac aggctccaaa
     1261 agttgataac aagtttaaca aggagcagca gaatgctttc tacgagatat tgcaccttcc
     1321 taatctgaat gaggaacaaa gaaatgcatt tatccagtcc ctaaaagacg atccatcaca
     1381 gagtgccaac ctgcttgcgg aagccaaaaa gctcaatgat gcccaggccc cgaaagtaga
     1441 cgcgaaccat caataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     1501 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1561 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1621 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1681 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     1741 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     1801 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     1861 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     1921 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     1981 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     2041 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     2101 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     2161 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     2221 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2281 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2341 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2401 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2461 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2521 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2581 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2641 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2701 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2761 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2821 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2881 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2941 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3001 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3061 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3121 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     3181 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     3241 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     3301 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     3361 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     3421 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     3481 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     3541 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     3601 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     3661 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3721 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     3781 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     3841 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3901 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     3961 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4021 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     4081 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4141 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     4201 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     4261 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4321 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4381 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4441 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     4501 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     4561 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     4621 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     4681 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     4741 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     4801 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     4861 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     4921 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     4981 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     5041 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5101 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5161 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5221 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5281 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5341 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     5401 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     5461 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     5521 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     5581 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
