LOCUS       sequence-317673-        4514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MelA in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4514)
  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 4514)
  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: enz.105-MelA
FEATURES             Location/Qualifiers
     source          1..4514
                     /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"
     misc_RNA        34..83
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             130..147
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             160..1257
                     /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
                     AYPIAVEALSLIYNKDLLPNQPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1114..1123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1231..1254
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     terminator      2153..2224
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2240..2267
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2312..2329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2483..3071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2563..2582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3230..4045)
                     /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(3230..4045)
                     /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     3953..3972
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4037..4056
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        complement(4147..4251)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4319..4337
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4403..4422
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agatactaga gagctgtcaa aggatgtgct ttccggtctg
       61 atgagtccgt gaggacgaaa cagcctctac aaataatttt gtttaagaaa gaggggacaa
      121 actagtatgc atcaccatca ccatcacggc ggtagtggca aaatcgaaga aggtaaactg
      181 gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc
      241 gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc
      301 ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt
      361 ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac
      421 aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg
      481 atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa ccagccaaaa
      541 acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg
      601 atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat
      661 gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc
      721 gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac
      781 accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac
      841 ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg
      901 ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac
      961 gccgccagtc cgaacaaaga gctggcgaaa gagttcctcg aaaactatct gctgactgat
     1021 gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac
     1081 gaggaagagt tggcgaaaga tccacgtatt gccgccacca tggaaaacgc ccagaaaggt
     1141 gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg
     1201 atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactcgt
     1261 atcaccaagg aactggttgc aagtgctaaa gaaaacctgt attttcaaga catgacagtc
     1321 cgtaaaaatc aagcatcctt gacggcggaa gagaaacgtc gttttgtagc ggcattactt
     1381 gaattaaaac gtaccggccg ttatgacgcg ttcgtgacta cccacaatgc attcattctg
     1441 ggggataccg ataacggtga acgcacaggc caccgctcgc ccagctttct gccctggcat
     1501 cgccgcttct tgttggagtt tgaacgcgca ctgcaatcgg tcgatgcgag cgtcgccctg
     1561 ccgtattggg attggtccgc ggatcgctca acccgctctt cgctgtgggc gccagatttt
     1621 ttgggcggca cgggccgtag ccgcgacggc caggttatgg atggcccttt cgcagcctcg
     1681 gcaggtaatt ggccgattaa tgtccgtgtg gacggccgca cgttcctccg ccgtgcgtta
     1741 ggcgccggcg tgagcgaact gccaacccgc gctgaagtgg acagcgtcct ggccatggct
     1801 acctacgata tggcgccctg gaactccggc agcgatggct ttcgcaatca tttagagggt
     1861 tggcgcggtg tcaatctgca taaccgcgtg cacgtttggg ttggtggtca aatggccacg
     1921 ggcgttagcc ccaatgatcc ggttttctgg ttacaccatg cctatattga taaattatgg
     1981 gcggaatggc agcgccgtca tccgagcagc ccgtatttgc caggcggagg tacgcccaac
     2041 gtggtggact taaatgaaac tatgaagccg tggaatgaca cgacccccgc agccctgtta
     2101 gatcataccc gtcactatac ctttgatgtc tgataatact agagccaggc atcaaataaa
     2161 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2221 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt
     2281 agcgtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2341 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2401 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2461 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2521 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2581 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2641 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2701 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2761 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2821 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2881 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2941 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3001 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3061 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ggtctgacgc tcagtggaac
     3181 gaaaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacct tagaaaaact
     3241 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     3301 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     3361 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     3421 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     3481 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     3541 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     3601 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     3661 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     3721 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     3781 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     3841 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     3901 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3961 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag
     4021 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     4081 gttttattgt tcatgatgat atatttttat cttgtgcaat gtacatcaga gattttgaga
     4141 cacaacactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4261 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4321 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4381 acacatgcag ctcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     4441 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgaattcgc
     4501 tgcagcttct agag
//
