LOCUS       sequence-311993-        4884 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 4884)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4884)
  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-m4133138
FEATURES             Location/Qualifiers
     source          1..4884
                     /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..1278
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1331..1452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1342..1361)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1396..1415
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1567..1613
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1643..1779
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1824..1895
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2121..2709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2201..2220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3455..3523,3524..4315)
                     /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(3673..3692)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4328..4464
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4616..4687
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4692..4738)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4756..4804
                     /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 aagaggcgat ggaaaaagaa ttttatcgcg gtgagtgcag ccaacagatt caaaaagata
     1261 tcctcgtctg gtgccctcta agctagcttg actgactgag atacagcgta ccttcagctc
     1321 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1381 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1441 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1501 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
     1561 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
     1621 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     1681 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     1741 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
     1801 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     1861 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     1921 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
     1981 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
     2041 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2521 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2941 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     3001 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     3061 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     3121 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     3181 cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     3241 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     3301 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     3361 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     3421 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     3481 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3541 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     3601 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     3661 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3721 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     3781 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3841 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     3901 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3961 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     4021 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     4081 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4141 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4201 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4261 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     4321 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     4381 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     4441 tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc tggctctatc
     4501 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     4561 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     4621 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     4681 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     4741 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     4801 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     4861 tcatagccca tatatggagt tccg
//
