LOCUS       sequence-339570-        6725 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 6725)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6725)
  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-m4134402
FEATURES             Location/Qualifiers
     source          1..6725
                     /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             1195..1212
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1265..1386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1276..1295)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1330..1349
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1501..1547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1577..1713
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1758..1829
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2055..2643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2135..2154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3143..4352
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4189..4208
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4356..4955
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4356..4375)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4852..4872
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4982..5030
                     /note="synthetic polyadenylation signal"
     CDS             join(5296..5364,5365..6156)
                     /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(5514..5533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6169..6305
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6457..6528
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6533..6579)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6597..6645
                     /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 gtcgcaccat
     1201 caccatcacc attaagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     1261 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1321 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1381 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1441 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     1501 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     1561 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     1621 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     1681 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     1741 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     1801 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     1861 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     1921 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     1981 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2461 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2701 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2761 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2821 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     2881 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     2941 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     3001 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     3061 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     3121 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     3181 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
     3241 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     3301 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     3361 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     3421 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     3481 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     3541 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     3601 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     3661 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     3721 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     3781 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     3841 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3901 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     3961 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     4021 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     4081 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     4141 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     4201 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     4261 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
     4321 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
     4381 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4441 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4501 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4561 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4621 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4681 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4741 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4801 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4861 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4921 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
     4981 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     5041 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     5101 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     5161 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     5221 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     5281 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5341 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5401 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5461 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     5521 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5581 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     5641 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5701 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5761 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5821 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     5881 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     5941 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6001 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6061 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6121 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     6181 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     6241 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     6301 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6361 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     6421 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     6481 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     6541 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     6601 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     6661 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     6721 ttccg
//
