LOCUS       sequence-333793-        6340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SARS-CoV-2 Nucleocapsid Protein expression in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6340)
  AUTHORS   Kolesov DE, Sinegubova MV, Safenkova IV, Vorobiev II, Orlova NA
  TITLE     Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by 
            the RNA admixture.
  JOURNAL   PeerJ. 2022 Jan 7;10:e12751. doi: 10.7717/peerj.12751. eCollection 
            2022.
  PUBMED    35036106
REFERENCE   2  (bases 1 to 6340)
  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: "PeerJ."; 
            date: "2022-01-7"; volume: "10:e12751. doi"; pages: " 
            10.7717/peerj.12751. eCollection 2022"
COMMENT     Sequence Label: pHYP-NPC-10H
FEATURES             Location/Qualifiers
     source          1..6340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     921..940
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(960..1037)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1243..1262
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1346..1365
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1346..1364
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1365..1389
                     /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             1404..1426
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2690..2719
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2745..2762
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2815..2833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2829..2876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2913..3271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3000..3019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3210..3231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4055..4870)
                     /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(4055..4870)
                     /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     4778..4797
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4862..4881
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4992..5580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5481..5500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5734..5751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5831..5850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(6217..6340,1..959))
                     /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"
ORIGIN
        1 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
       61 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
      121 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
      181 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
      241 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
      301 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
      361 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
      421 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
      481 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
      541 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
      601 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
      661 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
      721 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
      781 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
      841 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
      901 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
      961 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     1021 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     1081 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     1141 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     1201 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     1261 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     1321 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
     1381 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata ccatgtcaga
     1441 caatggtcct cagaaccagc gtaatgctcc taggatcact ttcggcggtc caagtgattc
     1501 gaccggcagc aatcaaaatg gtgagcgatc cggggcgcgt agcaagcagc gccgccctca
     1561 aggtctcccg aacaatacag cgtcttggtt cactgcgctg acgcagcatg gaaaggagga
     1621 tcttaagttt ccgcgtgggc agggcgtgcc gatcaacaca aacagttccc cagatgatca
     1681 gataggctac taccggcgcg cgacgcgacg aatacgcggc ggagacggca agatgaagga
     1741 cctgagtccg cggtggtact tttactatct gggcacaggg ccagaagcgg gcctaccgta
     1801 cggcgccaac aaagatggca tcatatgggt agcgacagaa ggggccctga acacccctaa
     1861 ggatcatatt ggcacaagga accctgccaa caatgctgcc atcgtgcttc aactcccgca
     1921 aggtactact ttgccgaagg gcttctacgc agagggttcg cgcggtggct cacaggcgag
     1981 ttcacgcagt agtagtcggt cccggaattc atcccgaaac tcgacacccg gctctagccg
     2041 tggtactagc ccagctagaa tggcaggcaa tggaggggat gccgcgctag ccttgctgct
     2101 cctcgataga ctcaaccagc tcgaaagcaa gatgtcagga aagggccagc agcagcaagg
     2161 acagactgtg acgaagaaaa gcgcggctga agcgagcaag aagccaagac aaaagcgcac
     2221 cgcaacaaag gcgtacaatg ttacacaggc ttttggaaga cgcggtccgg aacaaacgca
     2281 gggtaatttt ggagaccaag agttaattag gcagggtacg gactacaagc attggcctca
     2341 gatcgcccaa tttgccccgt cggcgtccgc gttctttggg atgtcgcgca ttggaatgga
     2401 agttaccccg agcggcacct ggttaacata caccggtgca attaagctag atgataaaga
     2461 tccaaacttc aaagaccaag taattctact caacaagcac atcgatgcct acaagacttt
     2521 tcctccaaca gagcctaaga aggacaagaa gaagaaagct gacgaaactc aggcactacc
     2581 ccaacgtcaa aagaagcaac agacagtcac actccttccc gccgcggatt tagatgattt
     2641 ctcgaaacag ctgcaacagt caatgtctag cgcggatagt acacaggcgc atcaccatca
     2701 tcaccaccat caccatcact gataagcttg cggccgcact cgagcaccac caccaccacc
     2761 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     2821 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2881 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     2941 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3001 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3061 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3121 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3181 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3241 caaccctatc tcggtctatt cttttgattt agtgcatgca caacatcagc aaggagaaag
     3301 gggctaccgg cgaaccagca gcccctttat aaaggcgctt cagtagtcag accagcatca
     3361 gtcctgaaaa ggcgggcctg cgcccgcctc caggttgcta cttaccggat tcgtaagcca
     3421 tgaaagccgc cacctccctg tgtccgtctc tgtaacgaat ctcgcacagc gattttcgtg
     3481 tcagataagt gaatatcaac agtgtgagac acacgatcaa cacacaccag acaagggaac
     3541 ttgtggtagt ttcatggcct tcttctcctt gcgcaaagcg cggtaagagg ctatcctgat
     3601 gtggactaga catagggatg cctcgtggtg gttaatgaaa attaacttac tacggggcta
     3661 tcttctttct gccacacaac acggcaacaa accaccttca cgtcatgagg cagaaagcct
     3721 caagcgccgg gcacatcata gcccatatac ctgcacgctg accacactca ctttccctga
     3781 aaataatccg ctcattcaga ccgttcacgg gaaatccgtg tgattgttgc cgcatcacgc
     3841 tgcctcccgg agtttgttgg atccactaag ggattttgcc gatttcggcc tattggttaa
     3901 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3961 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat
     4081 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     4141 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     4201 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     4261 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
     4321 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     4381 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     4441 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     4501 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     4561 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     4621 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     4681 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     4741 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     4801 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa
     4861 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     4921 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4981 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5041 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5101 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5161 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5221 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5281 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5761 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     5821 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtagcac ccgtggggcc
     5881 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     5941 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     6001 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     6061 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     6121 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     6181 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6241 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6301 attgggcgcc agggtggttt ttcttttcac cagtgagacg
//
