LOCUS       sequence-295273-        5094 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5094)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5094)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134108
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             872..892
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             893..910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    963..1084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(974..993)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1028..1047
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1199..1245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1275..1411
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1456..1527
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1753..2341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1833..1852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3087..3155,3156..3947)
                     /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(3305..3324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3960..4096
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4248..4319
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4324..4370)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4388..4436
                     /note="synthetic polyadenylation signal"
     enhancer        4517..4820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4822..5024
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4975..4995
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5093..5094,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gtgaaaagaa caatcaaggg tacacaccac tggttgttac tcacaatttt
       61 gacttcactt ttagttttag tccagagtac tcaatggtct ttgttctttt ttttgtatga
      121 aaatgccttt ttaccttttg ctatgggtat tattgctatg tctgcttttg caatgatgtt
      181 tgtcaaacat aagcatgcat ttctctgttt gtttttgtta ccttctcttg ccactgtagc
      241 ttattttaat atggtctata tgcctgctag ttgggtgatg cgtattatga catggttgga
      301 tatggttgat actagtttgt ctggttttaa gctaaaagac tgtgttatgt atgcatcagc
      361 tgtagtgtta ctaatcctta tgacagcaag aactgtgtat gatgatggtg ctaggagagt
      421 gtggacactt atgaatgtct tgacactcgt ttataaagtt tattatggta atgctttaga
      481 tcaagccatt tccatgtggg ctcttataat ctctgttact tctaactact caggtgtagt
      541 tacaactgtc atgtttttgg ccagaggtat tgtttttatg tgtgttgagt attgccctat
      601 tttcttcata actggtaata cacttcagtg tataatgcta gtttattgtt tcttaggcta
      661 tttttgtact tgttactttg gcctcttttg tttactcaac cgctacttta gactgactct
      721 tggtgtttat gattacttag tttctacaca ggagtttaga tatatgaatt cacagggact
      781 actcccaccc aagaatagca tagatgcctt caaactcaac attaaattgt tgggtgttgg
      841 tggcaaacct tgtatcaaag tagccactgt agaaaacctt tactttcagt cgcaccatca
      901 ccatcaccat taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      961 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1021 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1081 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1141 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     1201 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     1261 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1321 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     1381 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     1441 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1501 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1561 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1621 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1681 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2581 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2641 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2701 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2761 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2821 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     2881 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     2941 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     3001 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     3061 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     3121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3181 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     3241 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     3301 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3361 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     3421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3481 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     3541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     3661 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     3721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     3961 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     4021 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     4081 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     4141 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     4201 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     4261 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     4321 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     4381 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     4441 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     4501 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4561 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4621 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4681 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4741 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4801 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4861 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4921 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4981 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     5041 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
