LOCUS       sequence-334259-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of SARS-CoV-2 nucleocapsid protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  TITLE     Esposito Lab Plasmids
REFERENCE   2  (bases 1 to 6936)
  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: R619-X67-527: His6-SARS-CoV-2 N
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(9..33)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             93..125
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             132..149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      315..401
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      493..520
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(574..592)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      588..635
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      672..1127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(759..778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     969..990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1242..2033,2034..2102))
                     /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     1865..1884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2103..2195)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2276..2864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2765..2784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3018..3035
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3050..3189
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3115..3134)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3275..3297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3291..3482)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4054..5136)
                     /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     5098..5117
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5137..5214)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5420..5439
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5527..5546
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5527..5545
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5546..5570
                     /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             5585..5607
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5630..5647
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    5660..5684
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5873..5884)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 ggcttaatac ccaactttct tgtacaaagt ggtgatatcg ctagctagat agctagaagc
       61 ttgcggccgc acagctgtat acacgtgcaa gccagccaga actcgctcct gaagacccag
      121 aggatctcga gcaccaccac caccaccact aatgttaatt aagttgggcg ttcctaggct
      181 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
      241 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
      301 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      361 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      421 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
      481 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
      541 tgtttatttt tctaaataca ttcaaatatg tatccgctga gcaataacta gcataacccc
      601 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
      661 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      721 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      781 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      841 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      901 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      961 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1021 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1081 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
     1141 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1381 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2101 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
     2161 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
     2221 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2281 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2401 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     2461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2941 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3001 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3061 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     3121 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3181 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3241 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3301 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     3361 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     3421 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     3481 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     3541 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     3601 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     3661 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     3721 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3781 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3841 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3901 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac
     3961 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4021 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4141 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4201 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4261 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4321 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4381 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4441 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4501 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4561 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4621 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4681 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4741 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4801 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     4861 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     4921 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     4981 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5041 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5101 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5161 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5221 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5281 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5341 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5401 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5461 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     5521 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     5581 taattttgtt taactttaag aaggagatat acatatgaga tctggatccc accatcacca
     5641 tcaccataga tctgatatca caagtttgta caaaaaagtt ggcatgtccg acaacggtcc
     5701 ccagaaccag cgtaacgctc caaggatcac cttcggtgga cctagcgact ccaccggcag
     5761 caaccagaac ggagaacgct ccggagccag aagcaagcag cgtaggccac agggcctgcc
     5821 taacaacacc gccagctggt tcaccgctct gacccagcac ggaaaggaag acctgaagtt
     5881 cccaagggga cagggcgtgc caatcaacac caacagctct ccagacgacc agatcggata
     5941 ctacaggcgt gccaccaggc gcatcagagg aggcgacgga aagatgaagg acctgtctcc
     6001 taggtggtac ttctactacc tgggcaccgg accagaggct ggtctgccat acggcgccaa
     6061 caaggacggc atcatctggg tggctaccga gggagctctg aacaccccaa aggaccacat
     6121 cggaacccgt aaccctgcca acaacgctgc catcgtgctg cagctgccac agggaaccac
     6181 cctgcccaag ggattctacg ctgagggctc caggggtgga agccaggcta gctctcgttc
     6241 ttccagccgt tctaggaaca gctcccgcaa ctctacccca ggaagctcca gaggcaccag
     6301 cccagctcgt atggccggca acggaggtga cgccgctctg gctctgctgc tgctggaccg
     6361 tctgaaccag ctggaaagca agatgtccgg aaagggtcag cagcagcagg gccagaccgt
     6421 gaccaagaag tctgccgctg aggcctctaa gaagcccagg cagaagagga ccgctaccaa
     6481 ggcctacaac gtgacccagg ccttcggaag acgtggacct gaacagaccc agggaaactt
     6541 cggcgaccag gagctgatcc gccagggaac cgactacaag cactggccac agatcgctca
     6601 gttcgcccct tctgctagcg ccttcttcgg aatgtccagg atcggaatgg aggtgacccc
     6661 ttccggcacc tggctgacct acaccggagc tatcaagctg gacgacaagg accccaactt
     6721 caaggaccag gtgatcctgc tgaacaagca catcgacgcc tacaagacct tcccacctac
     6781 cgagccaaag aaggacaaga agaagaaggc cgacgaaacc caggccctgc cacagaggca
     6841 gaagaagcag cagaccgtga ccctgctgcc agctgctgac ctggacgact tctccaagca
     6901 gctgcagcag tctatgagct ctgctgactc caccca
//
