LOCUS       sequence-331339-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Nucleocapsid protein (residues 1-419).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  TITLE     Savchenko Lab Plasmids
REFERENCE   2  (bases 1 to 6065)
  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: pMCSG53_N
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     892..911
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(931..1008)
                     /gene="lacI"
                     /label=lacI promoter
     tRNA            1148..1223
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     primer_bind     1372..1391
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     tRNA            1493..1569
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     primer_bind     1844..1863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1844..1862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1863..1887
                     /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             1902..1924
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1935..1952
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1977..1997
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3323..3340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3393..3411)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3407..3454
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3578..3597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3764..3867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3868..3936,3937..4728)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4086..4105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4899..5487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5388..5407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5641..5658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5738..5757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(5913..6065,1..930))
                     /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 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
       61 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
      121 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
      181 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
      241 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
      301 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
      361 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
      421 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
      481 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
      541 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
      601 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
      661 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
      721 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
      781 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
      841 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
      901 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
      961 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     1021 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     1081 gcaccgccgc cgcaaggaat ggtgcatgcg gattgcgaca cggagttact ttataatccg
     1141 ctaccatggc cccttagctc agtggttaga gcaggcgact cataatcgct tggtcgctgg
     1201 ttcaagtcca gcaggggcca ccagatatag caaaggctga cgagaaatcg tcagcctttt
     1261 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     1321 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     1381 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggcg agatcaaaag ccattgactc
     1441 agcaagggtt gaccgtataa ttcacgcgat tacaccgcat tgcggtatca acgcgccctt
     1501 agctcagttg gatagagcaa cgaccttcta agtcgtgggc cgcaggttcg aatcctgcag
     1561 ggcgcgccat tacaattcaa tcagttacgc cttctttata tcctccataa ttccagagtg
     1621 ggacatattt gggacattat caccaaaaat gtcgtctatt ttcctcgcat gctctgtcaa
     1681 atgattaggc gcaaggtgag catacctacg aaccatttct atggactccc atccgcccat
     1741 ttcctgaagc actgataatg ggacgcctga ctgaatccac cggtgatgcc ggccacgatg
     1801 cgtccggcgt agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata
     1861 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     1921 gagatataca tatgcaccat catcatcatc attcttctgg tgtagatctg ggtaccgaga
     1981 acctgtactt ccaatccaat atgagcgata atggaccaca gaaccagcgt aatgcacctc
     2041 gtatcacctt tggtggtcct agtgatagta caggttcaaa tcagaacggt gaacgtagtg
     2101 gtgcacgcag taaacaaaga cgtcctcaag gtctgccgaa taatacagca agttggttta
     2161 cagcactgac ccagcacggt aaagaagacc tgaaatttcc gcgtggtcaa ggtgttccga
     2221 ttaataccaa tagtagcccg gacgaccaaa ttggctatta tagacgtgca acccgtcgca
     2281 ttcgtggtgg tgatggtaaa atgaaagacc tgtctcctcg ctggtatttc tattatctgg
     2341 gtaccggtcc tgaagcaggt ctgccttatg gtgcaaacaa agatggcatc atttgggttg
     2401 caacagaagg tgcactgaat accccgaaag accatattgg tacccgtaat ccggccaata
     2461 atgcagccat tgttttacag ctgcctcaag gtacaacact gcctaaaggc ttctatgcgg
     2521 aaggttcaag aggtggtagt caagcaagta gtcgtagttc aagtcgtagc cgtaatagca
     2581 gtcgcaatag tacacctggt agtagtcgtg gtacaagtcc tgcaagaatg gcaggtaatg
     2641 gtggtgatgc agcattagcc ttactgttac tggatcgcct gaatcaactg gagagcaaaa
     2701 tgagcggtaa aggtcagcag caacaaggtc agacagttac caagaaatca gcagcagaag
     2761 ccagcaaaaa acctcgtcag aaacgtacag ccaccaaagc ctataacgtt acacaggcat
     2821 ttggtcgtcg tggtccagaa caaacacaag gcaatttcgg cgatcaggaa ctgattagac
     2881 aaggtaccga ctacaaacat tggccgcaga ttgcacagtt tgcaccttca gcaagcgcgt
     2941 tttttggtat gagccgtatt ggtatggaag ttaccccttc aggtacatgg ctgacctata
     3001 caggcgccat taaactggac gacaaagatc cgaacttcaa agatcaggtg attctgctga
     3061 acaagcacat cgacgcgtac aagacctttc ctcctacaga gccgaaaaaa gacaagaaga
     3121 agaaagccga tgaaacacag gcactgcctc aacgtcagaa gaaacagcag accgttacct
     3181 tattacctgc agccgattta gatgatttca gcaagcagct gcagcagtca atgtcaagcg
     3241 cagatagtac acaagcataa attggaagtg gataacggat ccgaattcga gctccgtcga
     3301 caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     3361 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3421 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     3481 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3541 agagtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3721 cattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3841 gcttcaataa tttgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3901 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3961 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4021 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4081 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4141 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4201 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4261 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4321 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4381 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4441 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4501 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4561 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4621 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4681 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4741 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4801 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5041 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5281 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5521 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5581 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5641 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     5701 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     5761 agttaagcca gtatacactc cgctatcgct acgtgactgt agtcatgccc cgcgcccacc
     5821 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     5881 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5941 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6001 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     6061 gcctg
//
