LOCUS       sequence-313290-        4851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4851)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4851)
  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-m4133156
FEATURES             Location/Qualifiers
     source          1..4851
                     /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"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1180..1200
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1201..1245
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1298..1419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1309..1328)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1363..1382
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1534..1580
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1610..1746
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1791..1862
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2088..2676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2168..2187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3422..3490,3491..4282)
                     /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(3640..3659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4295..4431
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4583..4654
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4659..4705)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4723..4771
                     /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 ccatggcaat tgcctcagaa
      601 tttagttccc tcccctcgta cgctgcgttc gctaccgccc aagaggcata tgagcaggcc
      661 gtagccaacg gggacagcga agtggttctt aagaaactga agaaatcttt gaatgtggca
      721 aagagcgagt ttgatcggga cgcggcaatg cagaggaagc tggaaaaaat ggctgatcag
      781 gctatgacac agatgtacaa acaagcccga tccgaggaca agagagctaa agtcacttct
      841 gccatgcaaa cgatgctatt caccatgtta cgtaagctgg ataacgacgc actcaataac
      901 ataatcaata acgcccgcga tggttgtgtg ccattgaata tcattcctct tactacagcg
      961 gccaagctga tggtcgttat cccggactat aacacataca aaaatacctg cgacggcacc
     1021 actttcacgt atgctagcgc cctgtgggaa atacagcagg tagtggatgc cgattcaaag
     1081 attgtgcagc ttagtgagat ctccatggac aacagtccca acttggcttg gcctttaatc
     1141 gtcacagctc tcagagccaa tagcgcagtt aaactgcaag aaaaccttta ctttcagtcg
     1201 aaagaaaccg cggcagctaa gttcgagagg cagcacatgg acagttaagc tagcttgact
     1261 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     1321 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1381 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1441 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1501 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     1561 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     1621 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     1681 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     1741 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     1801 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     1861 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     1921 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     1981 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     2041 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2101 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2161 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2221 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2281 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2341 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2401 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2461 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2521 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2581 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2641 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2701 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2761 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2821 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2881 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     2941 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     3001 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     3061 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     3121 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     3181 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     3241 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     3301 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     3361 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     3421 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3481 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3541 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3601 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3661 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3721 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3781 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3841 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3901 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3961 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     4021 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4081 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4141 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4201 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4261 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     4321 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     4381 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     4441 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     4501 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     4561 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     4621 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     4681 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     4741 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     4801 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
