LOCUS       sequence-316813-        5635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NMT1 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5635)
  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 5635)
  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.083-NMT1
FEATURES             Location/Qualifiers
     source          1..5635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             18..35
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             48..1145
                     /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      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1119..1142
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1179..1199
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2707..2778
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2794..2821
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2841..2898
                     /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(3056..3871)
                     /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(3056..3871)
                     /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     3779..3798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3863..3882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(4466..5011)
                     /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(4546..4565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5524..5543
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5524..5542
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        5557..5606
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 ggggacaaac tagtatgcat caccatcacc atcacggcgg tagtggcaaa atcgaagaag
       61 gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta
      121 agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag
      181 agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg
      241 accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt
      301 tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg
      361 cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc
      421 agccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga
      481 gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg
      541 ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata
      601 acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga
      661 atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga
      721 ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa
      781 cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag
      841 gtattaacgc cgccagtccg aacaaagagc tggcgaaaga gttcctcgaa aactatctgc
      901 tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga
      961 agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg gaaaacgccc
     1021 agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta
     1081 ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc
     1141 agactcgtat caccaaggaa ctggttgcaa gtgctaaaga aaacctgtat tttcagggcg
     1201 cagacgagag tgaaactgcc gtcaaaccgc ctgctccacc gttgccccag atgatggagg
     1261 gtaatgggaa tggtcatgaa cattgttcag attgcgagaa tgaagaggat aactcgtata
     1321 accggggcgg actctcccca gctaacgata ccggggcgaa gaaaaagaag aagaaacaga
     1381 agaaaaagaa ggaaaaaggc tcggaaacag atagcgccca agaccaaccg gtaaaaatga
     1441 actcgttacc ggcagaacgg atccaagaaa tacagaaagc gattgaactg ttcagtgtcg
     1501 ggcagggccc ggctaaaacc atggaagagg cgagcaaacg ctcgtatcag ttttgggaca
     1561 cacagccggt ccctaagctg ggtgaagtag tgaatacgca cggtcccgtg gaaccggata
     1621 aagataatat ccgtcaggaa ccgtataccc tgccccaggg tttcacatgg gacgctctgg
     1681 acctggggga tcgtggagtc ctcaaagaat tgtacaccct cttaaacgaa aactatgttg
     1741 aagatgatga taatatgttc cgcttcgatt acagtccgga atttctactg tgggccctgc
     1801 gccctccagg ttggcttccg caatggcatt gtggcgtccg ggtagtgtcg tcccgtaaat
     1861 tggtcggttt tattagtgcg attccggcga atatccatat ttacgatacc gaaaaaaaga
     1921 tggttgaaat caacttttta tgtgttcaca agaagctgag aagcaaacgc gtagccccgg
     1981 tgctgatccg agaaattaca cgccgggtgc atctggaggg tatttttcag gcggtttata
     2041 cagcgggcgt cgtactgcca aaacccgtgg gcacttgccg ttattggcat cggagtctca
     2101 acccacgtaa acttattgag gtgaaattct cccatctgtc ccgcaatatg actatgcagc
     2161 gtacgatgaa attgtaccgt cttcccgaaa ccccaaaaac cgcaggcctg cgcccaatgg
     2221 aaacgaagga tatcccggtc gtacatcagc tgttaacccg ctatctgaag cagtttcatc
     2281 tgacaccggt gatgtcacag gaagaagtgg aacattggtt ctatccgcag gagaacatca
     2341 ttgatacatt tgtggtcgaa aatgcaaacg gtgaagttac cgatttctta tcgttttata
     2401 ctctgccgtc aacgatcatg aatcacccga cccataaatc actgaaagcc gcatacagct
     2461 tctacaatgt gcatacgcaa actcccctgt tagatctgat gagtgacgcg ctagtactgg
     2521 cgaaaatgaa gggttttgat gtgttcaacg cgctggatct gatggaaaat aaaaccttcc
     2581 ttgaaaagct gaaattcggt attggtgacg gcaatctgca gtattacctg tataattgga
     2641 aatgtccgtc aatgggagca gaaaaggtcg gcttggtact gcaataataa tactagagcc
     2701 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2761 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     2821 gtttatatac tagtagccag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
     2881 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
     2941 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatctcgagt
     3001 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     3061 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     3121 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     3181 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     3241 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     3301 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3361 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     3421 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     3481 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     3541 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3601 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3661 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3721 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     3781 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg
     3841 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     3901 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     3961 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     4021 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     4081 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     4141 atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcagc
     4201 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     4261 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     4321 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     4381 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     4441 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     4501 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     4561 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     4621 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     4681 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     4741 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     4801 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     4861 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     4921 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     4981 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg ggtctgacgc
     5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5401 atccgcctcc atccagtcta ttccatggtg ccacctgacg tctaagaaac cattattatc
     5461 atgacattaa cctataaaaa taggtgtatc acgagtcgct tgaattcgct gcagcttcta
     5521 gagtaatacg actcactata gggagatact agagagctgt caaaggatgt gctttccggt
     5581 ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaag aaaga
//
