LOCUS       sequence-295370-        5184 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5184)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5184)
  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-m4134133
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..445
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             479..556
                     /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"
     CDS             584..604
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1053..1174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1064..1083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1118..1137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1289..1335
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1365..1501
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1546..1617
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1843..2431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1923..1942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3177..3245,3246..4037)
                     /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(3395..3414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4050..4186
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4338..4409
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4414..4460)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4478..4526
                     /note="synthetic polyadenylation signal"
     enhancer        4607..4910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4912..5114
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5065..5085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5183..5184,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gggaatgcga cagaggttcc tgccaactca acggtcttgt ctttttgtgc
       61 tttcgccgtg gacgctgcaa aagcctacaa ggattatcta gcctcgggtg gacagccaat
      121 cactaactgc gtaaagatgc tttgtaccca caccggcact ggccaagcaa ttacagtgac
      181 ccccgaagct aatatggacc aggagtcctt cggaggtgcg agctgctgcc tgtactgtcg
      241 ttgtcatata gatcacccga atcccaaagg gttttgcgac ctcaagggca aatatgtcca
      301 gatcccaacg acatgcgcca acgatcctgt gggattcact ctgaagaaca ccgtatgtac
      361 cgtttgcggg atgtggaagg ggtatggctg tagttgcgac caattacggg aaccgatgct
      421 gcaagaaaac ctttactttc agtcgggacg gcgaatcccg gggctgataa atccctggaa
      481 aaggagatgg aagaagaact ttattgcagt gtctgcggct aaccggttca aaaagatcag
      541 ttcaagcggc gccctagatt acgacattcc tacgaccgcc tccgagaatc tctatttcca
      601 gggtgaattt ggcttggcac aacatgacga ggccgtcgat aacaagttta ataaagaaca
      661 gcagaacgct ttctatgaga tccttcactt accaaatctg aacgaagagc aacgcaacgc
      721 tttcattcag agcctgaaag acgatccctc gcagtctgca aatctcctgg ccgaagccaa
      781 gaagctgaat gacgcacagg ctccaaaagt tgataacaag tttaacaagg agcagcagaa
      841 tgctttctac gagatattgc accttcctaa tctgaatgag gaacaaagaa atgcatttat
      901 ccagtcccta aaagacgatc catcacagag tgccaacctg cttgcggaag ccaaaaagct
      961 caatgatgcc caggccccga aagtagacgc gaaccatcaa taagctagct tgactgactg
     1021 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     1081 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     1141 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     1201 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1261 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     1321 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     1381 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     1441 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     1501 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     1561 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     1621 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     1681 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     1741 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     1801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2581 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2641 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     2701 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     2761 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     2821 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     2881 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     2941 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     3001 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     3061 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     3121 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     3181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     3301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3361 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3481 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     3601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3721 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     3781 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4021 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     4081 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     4141 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     4201 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     4261 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     4321 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     4381 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     4441 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     4501 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     4561 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4681 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4741 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4801 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4861 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     4921 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4981 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5041 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5101 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     5161 ctcgacacac ccgccagcgg ccgc
//
