LOCUS       sequence-342386-        7200 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 7200)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7200)
  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-m4134010
FEATURES             Location/Qualifiers
     source          1..7200
                     /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..1242
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1311..1432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1322..1341)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1376..1395
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1547..1593
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1623..1759
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1804..1875
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2101..2689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2181..2200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3189..4398
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4235..4254
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4402..5430
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5457..5505
                     /note="synthetic polyadenylation signal"
     CDS             join(5771..5839,5840..6631)
                     /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(5989..6008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6644..6780
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6932..7003
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7008..7054)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7072..7120
                     /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 gggaaaccta tacccaatcc acttctcggc ttagattcta cctaaggatc cgcaggcctc
     1261 tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     1321 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1381 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1441 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1501 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     1561 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     1621 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     1681 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     1741 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     1801 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     1861 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     1921 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1981 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     2041 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2281 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2521 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2581 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     2941 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     3001 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     3061 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     3121 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     3181 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     3241 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     3301 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     3361 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     3421 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     3481 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     3541 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     3601 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     3661 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     3721 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     3781 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     3841 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     3901 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     3961 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     4021 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     4081 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     4141 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     4201 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     4261 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     4321 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     4381 ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag
     4441 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     4501 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     4561 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc cgccgcgctc
     4621 ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc
     4681 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     4741 cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca gacgagcggg
     4801 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     4861 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     4921 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     4981 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     5041 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     5101 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     5161 catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat tggtcttgac
     5221 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     5281 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     5341 agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     5401 cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc ttagataata
     5461 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     5521 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     5581 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     5641 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     5701 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     5761 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5821 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5881 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5941 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6001 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6061 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     6121 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6181 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6241 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6301 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     6361 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     6421 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6481 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6541 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6601 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     6661 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     6721 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     6781 gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     6841 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     6901 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     6961 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     7021 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     7081 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     7141 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
