LOCUS       sequence-313279-        5151 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 5151)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5151)
  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-m4133317
FEATURES             Location/Qualifiers
     source          1..5151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1456..1476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1480..1545
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1598..1719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1609..1628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1663..1682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1834..1880
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1910..2046
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2091..2162
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2388..2976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2468..2487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3722..3790,3791..4582)
                     /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(3940..3959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4595..4731
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4883..4954
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4959..5005)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5023..5071
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagcgc cgttaagcgg
      601 acaatcaaag ggacccatca ctggttgctc cttactatac tgacgtcttt actggtcttg
      661 gtacagagta cacaatggtc gcttttcttt tttctgtacg agaatgcatt cctcccattt
      721 gcgatgggaa tcattgctat gtccgccttc gctatgatgt tcgtgaagca caaacatgcc
      781 ttcctttgcc tgtttctcct accctcactg gcaaccgtgg cctattttaa catggtgtac
      841 atgcctgcga gctgggtcat gcgaattatg acttggctgg acatggttga tacctcccta
      901 tctggcttca agttaaagga ctgtgtgatg tatgcatcag ctgtagtgct gctaatcctg
      961 atgacagctc gcacagtcta cgatgacggt gccaggagag tgtggactct gatgaatgtc
     1021 ctcaccttag tttataaagt atactatggc aacgctctcg atcaggccat tagtatgtgg
     1081 gcattgataa tctccgtgac gagtaactac agcggagtgg taaccactgt catgtttttg
     1141 gcacgcggga tagtgttcat gtgtgtcgaa tattgcccga tcttttttat cacagggaat
     1201 actctgcaat gcattatgct ggtttactgt ttcttgggct atttctgcac ctgttatttt
     1261 ggtctattct gcctgcttaa taggtacttt cgtttgacac taggagtcta tgattacttg
     1321 gtgagcacgc aggagtttcg gtatatgaac agtcagggcc tgcttccccc aaagaactct
     1381 attgacgcgt ttaaactcaa tataaagctg ttgggcgtgg gtggaaaacc ttgcatcaag
     1441 gtggccactg tgcaggaaaa cctttacttt cagtcgatgg attataagga ccacgatgga
     1501 gactacaaag atcatgacat tgattacaag gacgacgatg acaaataagc tagcttgact
     1561 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     1621 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1681 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1741 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1801 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     1861 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     1921 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     1981 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     2041 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     2101 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     2161 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     2221 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     2281 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     2341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2821 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3121 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3181 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     3241 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     3301 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     3361 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     3421 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     3481 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     3541 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     3601 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     3661 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     3721 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3841 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3901 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4021 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4141 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4261 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     4321 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4501 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4561 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     4621 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     4681 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     4741 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     4801 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     4861 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     4921 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     4981 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     5041 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     5101 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
