LOCUS       sequence-317670-        5467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GCN5 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5467)
  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 5467)
  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.079-GCN5
FEATURES             Location/Qualifiers
     source          1..5467
                     /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"
     CDS             1291..1311
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2651..2722
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2738..2765
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2785..2842
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     CDS             complement(3000..3815)
                     /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(3000..3815)
                     /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     3723..3742
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3807..3826
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(4410..4955)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4490..4509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward 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 attttcaggg catggtcaca
     1321 aaacatcaga ttgaagagga tcatctggac ggtgccacaa cggacccaga agtcaaacgc
     1381 gtcaagctgg aaaataacgt ggaagaaatt cagccggaac aggccgagac caataaacag
     1441 gaaggtacgg ataaagagaa caagggcaaa tttgagaaag aaacagaaag gattggcggc
     1501 agtgaagtcg ttacagatgt cgaaaaaggc atcgtgaaat tcgagttcga cggcgtagaa
     1561 tataccttca aggaacgccc ctctgtggtg gaggagaacg aggggaaaat cgagtttcgc
     1621 gttgtaaata atgataatac gaaagaaaac atgatggttc taactggcct gaaaaatatc
     1681 ttccagaaac aactgcccaa aatgcccaag gaatacattg cacgcctggt ctatgatcga
     1741 tcgcatcttt cgatggctgt tatccgcaaa cctttaactg tggtgggcgg cattacttac
     1801 cgtccgttcg ataaacgtga attcgcggaa atcgtcttct gcgcaatttc ctcaacggag
     1861 caagtgcgcg gttacggggc acatctgatg aaccatctga aagattatgt tcgcaatacc
     1921 agcaacatta aatatttcct gacgtatgca gataactatg ccattggtta tttcaaaaag
     1981 cagggcttta ccaaagaaat tacccttgat aaaagcattt ggatgggtta cattaaagat
     2041 tatgaaggtg gaaccctcat gcagtgcagc atgctgccgc gaattcgcta cctggacgct
     2101 ggtaaaattt tgctgctgca ggaagccgcc ttgcgtcgca aaatccgcac gatttcaaaa
     2161 tcgcacatcg tgcgcccagg actggaacaa tttaaagatc tgaataacat caaaccgata
     2221 gatccgatga ctattccggg attgaaagag gctgggtgga cacccgagat ggatgccctg
     2281 gcgcagaggc cgaagcgtgg ccctcacgat gcggcaatac agaatatctt gacggagctc
     2341 cagaaccatg cggccgcgtg gccgtttctg caaccggtga acaaggagga agttccagac
     2401 tactacgatt tcattaaaga gccgatggat ttaagcacta tggaaatcaa gctggaaagc
     2461 aataaatacc agaaaatgga agattttatc tatgatgcga gactggtatt caataactgc
     2521 cgtatgtaca atggcgaaaa cacttcatat tacaaatatg cgaaccgcct cgaaaaattt
     2581 ttcaacaata aggtgaaaga aattccggag tactcacacc tgatcgacta ataatactag
     2641 agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2701 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     2761 ctgcgtttat atactagtag ccagtccggc aaaaaagggc aaggtgtcac caccctgccc
     2821 tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc
     2881 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatctc
     2941 gagtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat
     3001 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     3061 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     3121 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     3181 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     3241 gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag
     3301 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     3361 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     3421 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     3481 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     3541 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     3601 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     3661 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     3721 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     3781 ctggagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     3841 taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag
     3901 agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat
     3961 cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc
     4021 accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg
     4081 gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct
     4141 cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     4201 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     4261 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     4321 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     4381 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     4441 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     4501 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     4561 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     4621 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     4681 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     4741 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     4801 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     4861 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     4921 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg
     4981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5341 ctttatccgc ctccatccag tctattccat ggtgccacct gacgtctaag aaaccattat
     5401 tatcatgaca ttaacctata aaaataggtg tatcacgagt cgcttgaatt cgctgcagct
     5461 tctagag
//
