LOCUS       sequence-313249-        5190 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 5190)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5190)
  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-m4133314
FEATURES             Location/Qualifiers
     source          1..5190
                     /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             1174..1194
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1543..1560
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1637..1758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1648..1667)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1702..1721
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1873..1919
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1949..2085
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2130..2201
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2427..3015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2507..2526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3761..3829,3830..4621)
                     /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(3979..3998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4634..4770
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4922..4993
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4998..5044)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5062..5110
                     /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 ccatgatagc ctcagagttt
      601 agttcccttc catcatatgc agcttttgct actgctcaag aagcttatga gcaggctgtt
      661 gctaatggtg attctgaagt tgttcttaaa aagttgaaga agtctttgaa tgtggctaaa
      721 tctgaatttg accgtgatgc agccatgcaa cgtaagttgg aaaagatggc tgatcaagct
      781 atgacccaaa tgtataaaca ggctagatct gaggacaaga gggcaaaagt tactagtgct
      841 atgcagacaa tgcttttcac tatgcttaga aagttggata atgatgcact caacaacatt
      901 atcaacaatg caagagatgg ttgtgttccc ttgaacataa tacctcttac aacagcagcc
      961 aaactaatgg ttgtcatacc agactataac acatataaaa atacgtgtga tggtacaaca
     1021 tttacttatg catcagcatt gtgggaaatc caacaggttg tagatgcaga tagtaaaatt
     1081 gttcaactta gtgaaattag tatggacaat tcacctaatt tagcatggcc tcttattgta
     1141 acagctttaa gggccaattc tgctgtcaaa ttagaaaacc tttactttca gtcgatgagt
     1201 gacaaaatca ttcatctaac cgatgactcc tttgatacag acgtgttgaa ggccgatggt
     1261 gcgatactgg tagatttctg ggctgaatgg tgtggcccgt gcaagatgat cgcacccatt
     1321 ctggacgaga tcgccgacga gtatcaagga aaattaactg tcgctaagct taatatagat
     1381 cagaacccag ggacggcacc taaatacggc atcaggggaa tacctaccct cctgctcttt
     1441 aagaatgggg aagttgccgc gacaaaagtg ggagccctgt ctaagggcca gctaaaagag
     1501 ttcttggacg ctaacctggc aggttcaggc tcggggcaca tgcaccatca ccatcaccat
     1561 agcagcgggc tcgtgccacg cggctaagct agcttgactg actgagatac agcgtacctt
     1621 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1681 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1741 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1801 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1861 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1921 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1981 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     2041 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     2101 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     2161 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     2221 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     2281 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     2341 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     2401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3121 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3181 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3241 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     3301 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     3361 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     3421 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     3481 gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga tcctctcagt
     3541 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     3601 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     3661 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     3721 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     3781 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3841 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     3901 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     3961 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4021 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     4081 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4141 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     4201 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4261 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4321 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     4381 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4441 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4501 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4561 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4621 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     4681 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     4741 gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc
     4801 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     4861 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     4921 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     4981 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     5041 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     5101 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     5161 attagttcat agcccatata tggagttccg
//
