LOCUS       sequence-307572-        4698 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 4698)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4698)
  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-m4134399
FEATURES             Location/Qualifiers
     source          1..4698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..445
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             449..514
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    567..688
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(578..597)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     632..651
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      803..849
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      879..1015
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1060..1131
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1357..1945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1437..1456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2691..2759,2760..3551)
                     /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(2909..2928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3564..3700
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3852..3923
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3928..3974)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3992..4040
                     /note="synthetic polyadenylation signal"
     enhancer        4121..4424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4426..4628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4579..4599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4697..4698,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gggaatgcga cagaggttcc tgccaactca acggtcttgt ctttttgtgc
       61 tttcgccgtg gacgctgcaa aagcctacaa ggattatcta gcctcgggtg gacagccaat
      121 cactaactgc gtaaagatgc tttgtaccca caccggcact ggccaagcaa ttacagtgac
      181 ccccgaagct aatatggacc aggagtcctt cggaggtgcg agctgctgcc tgtactgtcg
      241 ttgtcatata gatcacccga atcccaaagg gttttgcgac ctcaagggca aatatgtcca
      301 gatcccaacg acatgcgcca acgatcctgt gggattcact ctgaagaaca ccgtatgtac
      361 cgtttgcggg atgtggaagg ggtatggctg tagttgcgac caattacggg aaccgatgct
      421 gcaagaaaac ctttactttc agtcgatgga ttataaggac cacgatggag actacaaaga
      481 tcatgacatt gattacaagg acgacgatga caaataagct agcttgactg actgagatac
      541 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
      601 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      661 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
      721 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
      781 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
      841 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
      901 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
      961 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     1021 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     1081 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     1141 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     1201 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     1261 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     1321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1861 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1921 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1981 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2041 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2101 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2161 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     2221 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     2281 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     2341 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     2401 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     2461 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     2521 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     2581 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     2641 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     2701 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2761 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     2821 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     2881 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2941 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     3001 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3061 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     3121 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3181 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3241 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3301 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3361 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3421 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3481 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3541 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     3601 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     3661 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     3721 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     3781 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     3841 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     3901 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     3961 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     4021 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     4081 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4141 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4201 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4261 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4321 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4381 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     4441 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4501 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4561 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4621 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     4681 acacccgcca gcggccgc
//
