LOCUS       sequence-317698-        3653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses R5 peptide in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3653)
  AUTHORS   Wallace AK, Chanut N, Voigt CA
  TITLE     Silica Nanostructures Produced Using Diatom Peptides with Designed 
            Post-Translational Modifications
  JOURNAL   Advanced Functional Materials 2020
REFERENCE   2  (bases 1 to 3653)
  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: "Advanced 
            Functional Materials 2020"
COMMENT     Sequence Label: pAKW.031-GM-R5
FEATURES             Location/Qualifiers
     source          1..3653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     285..302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(405..452)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     448..466
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(530..550)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(563..580)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(602..1702)
                     /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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNQPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(605..628)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     regulatory      736..745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1762..1786
                     /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)."
     primer_bind     complement(1786..1805)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1787..1805)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1889..1908)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1974..1992)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2060..2164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2165..2233,2234..3025)
                     /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(2383..2402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3196..3653,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      301 aggaagcgga agagcgccca atacgcaaac cacacccgcc gcgcttaatg cgccgctaca
      361 gggcgcgtcc cattcgccaa tccggatata gttcctcctt tcagcaaaaa acccctcaag
      421 acccgtttag aggccccaag gggttatgct agttattgct cagcggtcat aagatacgac
      481 gtttactgcc tttagagccg gaatatgaac cagatttttt ggaggacatg ccctgaaaat
      541 acaggttttc gccactaccg ccgtgatggt gatggtgatg gtagtacgac ttggtgatac
      601 gagtctgcgc gtctttcagg gcttcatcga cagtctgacg accgctggcg gcgttgatca
      661 ccgcagtacg cacggcatac cagaaagcgg acatctgcgg gatgttcggc atgatttcac
      721 ctttctgggc gttttccatg gtggcggcaa tacgtggatc tttcgccaac tcttcctcgt
      781 aagacttcag cgctacggca cccagcggtt tgtctttatt aaccgcttcc agaccttcat
      841 cagtcagcag atagttttcg aggaactctt tcgccagctc tttgttcgga ctggcggcgt
      901 taatacctgc gctcagcacg ccaacgaacg gtttggatgg ttgacccttg aaggtcggca
      961 gtaccgttac accataattc actttgctgg tgtcgatgtt ggaccatgcc cacgggccgt
     1021 tgatggtcat cgctgtttcg cctttattaa aggcagcttc tgcgatggag taatcggtgt
     1081 ctgcattcat gtgtttgttt ttaatcaggt caaccaggaa ggtcagaccc gctttcgcgc
     1141 cagcgttatc cacgcccacg tctttaatgt cgtacttgcc gttttcatac ttgaacgcat
     1201 aacccccgtc agcagcaatc agcggccagg tgaagtacgg ttcttgcagg ttgaacatca
     1261 gcgcgctctt acctttcgct ttcagttctt tatccagcgc cgggatctct tcccaggttt
     1321 ttggctggtt cggcagcaga tctttgttat aaatcagcga taacgcttca acagcgatcg
     1381 ggtaagcaat cagcttgccg ttgtaacgta cggcatccca ggtaaacgga tacagcttgt
     1441 cctggaacgc tttgtccggg gtgatttcag ccaacaggcc agattgagcg tagccaccaa
     1501 agcggtcgtg tgcccagaag ataatgtcag ggccatcgcc agttgccgca acctgtggga
     1561 atttctcttc cagtttatcc ggatgctcaa cggtgacttt aattccggta tctttctcga
     1621 atttcttacc gacttcagcg agaccgttat agcctttatc gccgttaatc cagattacca
     1681 gtttaccttc ttcgattttc attattttaa ccgtattatt tatagttgag tcgcaaagtt
     1741 aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg
     1801 tattaatttc gcgggatcga gatctcgatc ctctacgccg gacgcatcgt ggccggcatc
     1861 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagag
     1921 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     1981 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2041 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2101 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2161 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2221 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2281 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2341 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2401 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2461 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2521 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2581 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2641 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2701 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2761 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2821 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2881 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2941 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3001 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3061 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3121 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3181 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3241 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3301 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3361 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3421 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcg
//
