LOCUS       sequence-314768-        7584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express MBP tagged SARS-CoV-2 Nucleocapsid protein without the 
            N-terminal IDR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7584)
  AUTHORS   Perdikari TM, Murthy AC, Ryan VH, Watters S, Naik MT, Fawzi NL
  TITLE     SARS-CoV-2 nucleocapsid protein phase-separates with RNA and with 
            human hnRNPs.
  JOURNAL   EMBO J. 2020 Nov 17:e106478. doi: 10.15252/embj.2020106478.
  PUBMED    33200826
REFERENCE   2  (bases 1 to 7584)
  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"; journalName: "EMBO J. 
            2020 Nov 17:e106478. doi: 10.15252/embj.2020106478."
COMMENT     Sequence Label: SARS-CoV-2_N_deltaNIDR
FEATURES             Location/Qualifiers
     source          1..7584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             106..123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             136..1233
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1090..1099
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1207..1230
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1252..1276
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1285..1305
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1769..1786)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             2449..2466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2519..2537)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2533..2580
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2617..3072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2704..2723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2914..2935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3164..3979)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3164..3979)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3887..3906
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3971..3990
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4101..4689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4590..4609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4843..4860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4875..5014
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4940..4959)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5100..5122
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5116..5307)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6116..7198)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7160..7179
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7199..7276)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7482..7501
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggatctgata aaattcatca tcatcatcat
      121 cacagcagcg gtaccaaaat cgaagaaggt aaactggtaa tctggattaa cggcgataaa
      181 ggctataacg gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc
      241 accgttgagc atccggataa actggaagag aaattcccac aggttgcggc aactggcgat
      301 ggccctgaca ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg
      361 ttggctgaaa tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat
      421 gccgtacgtt acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg
      481 atttataaca aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg
      541 gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac
      601 ttcacctggc cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag
      661 tacgacatta aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg
      721 gttgacctga ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct
      781 gcctttaata aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc
      841 gacaccagca aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc
      901 aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg
      961 gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa
     1021 gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca
     1081 cgtattgccg ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag
     1141 atgtccgctt tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag
     1201 actgtcgatg aagccctgaa agacgcgcag actaattcgg gatctgatat cacaagtttg
     1261 tacaaaaaag ctgaaggcgg taccgagaac ctgtacttcc agggtcatat ggtgctgccg
     1321 aacaacaccg cgagctggtt taccgcgctg acccaacacg gcaaagaaga cctgaaattc
     1381 ccgcgtggtc aaggcgttcc gatcaacacc aacagcagcc cggatgacca gattggctat
     1441 taccgtcgtg cgacccgtcg tattcgtggc ggtgatggca agatgaaaga cctgagcccg
     1501 cgttggtatt tctactatct gggtaccggc ccggaggcgg gtctgccgta tggtgcgaac
     1561 aaggacggta tcatttgggt ggcgaccgaa ggcgcgctga acaccccgaa ggaccacatc
     1621 ggtacccgta acccggcgaa caacgcggcg atcgttctgc agctgccgca aggtaccacc
     1681 ctgccgaaag gtttctatgc ggagggtagc cgtggcggta gccaagcgag cagccgtagc
     1741 agcagccgta gccgtaacag cagccgtaac agcaccccgg gcagcagccg tggtaccagc
     1801 ccggcgcgta tggcgggcaa cggtggtgac gcggcgctgg cgctgctgct gctggatcgt
     1861 ctgaaccaac tggagagcaa gatgagcggc aagggtcaac agcaacaagg tcaaaccgtt
     1921 accaagaaaa gcgcggcgga ggcgagcaaa aagccgcgtc agaaacgtac cgcgaccaaa
     1981 gcgtacaacg tgacccaagc gtttggtcgt cgtggtccgg aacaaaccca gggtaacttc
     2041 ggtgatcagg aactgattcg tcaaggtacc gactacaaac actggccgca aatcgcgcag
     2101 ttcgcgccga gcgcgagcgc gttctttggt atgagccgta tcggtatgga agttaccccg
     2161 agcggtacct ggctgaccta taccggtgcg atcaagctgg acgataagga tccgaacttc
     2221 aaggaccagg ttatcctgct gaacaagcac atcgacgcgt acaaaacctt cccgccgacc
     2281 gagccgaaga aagacaagaa gaagaaagcg gacgagaccc aagcgctgcc gcaacgtcag
     2341 aaaaagcaac agaccgttac cctgctgccg gcggcggatc tggacgactt tagcaaacaa
     2401 ctgcaacaga gcatgagcag cgcggatagc acccaggcgt aactcgagca ccaccaccac
     2461 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     2521 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2581 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     2641 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2701 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2761 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2821 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2881 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2941 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3001 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3061 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3121 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     3181 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     3241 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     3301 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     3361 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3421 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3481 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3541 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     3601 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     3661 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     3721 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     3781 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     3841 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     3901 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     3961 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     4021 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4081 gaaaagatca aaggatcttc ttgatatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4741 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4801 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4861 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4921 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     4981 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     5041 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5101 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     5161 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     5221 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     5281 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     5341 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     5401 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     5461 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     5521 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     5581 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     5641 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     5701 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     5761 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     5821 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     5881 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     5941 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     6001 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     6061 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     6121 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6181 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     6241 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     6301 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     6361 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     6421 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     6481 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     6541 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     6601 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     6661 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     6721 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     6781 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     6841 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     6901 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     6961 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     7021 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     7081 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     7141 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     7201 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     7261 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     7321 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     7381 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     7441 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     7501 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     7561 atcgagatct cgatcccgcg aaat
//
