LOCUS       sequence-317669-        5248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SETD7 (Y305F) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5248)
  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 5248)
  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.043-SETD7-Y305F
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     465..484
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        465..483
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        498..547
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             594..611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             624..1721
                     /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
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1578..1587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1695..1718
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1755..1775
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2896..2967
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2983..3010
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      3030..3087
                     /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(3245..4060)
                     /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
                     TDRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3968..3987
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4052..4071
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(4655..5200)
                     /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(4735..4754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
       61 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
      181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
      361 attccatggt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
      421 ataggtgtat cacgagtcgc ttgaattcgc tgcagcttct agagtaatac gactcactat
      481 agggagatac tagagagctg tcaaaggatg tgctttccgg tctgatgagt ccgtgaggac
      541 gaaacagcct ctacaaataa ttttgtttaa gaaagagggg acaaactagt atgcatcacc
      601 atcaccatca cggcggtagt ggcaaaatcg aagaaggtaa actggtaatc tggattaacg
      661 gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa
      721 ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa
      781 ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat
      841 ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta
      901 cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt
      961 tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc
     1021 cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag
     1081 aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa
     1141 acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga
     1201 ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg
     1261 cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt
     1321 ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc
     1381 aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca
     1441 aagagctggc gaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg
     1501 ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga
     1561 aagatccacg tattgccgcc accatggaaa acgcccagaa aggtgaaatc atgccgaaca
     1621 tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg
     1681 gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac tcgtatcacc aaggaactgg
     1741 ttgcaagtgc taaagaaaac ctgtattttc aaggcatgga tagcgacgac gagatggtgg
     1801 aggaggcggt ggaagggcac ctggacgatg acggattacc gcacgggttc tgcacagtca
     1861 cctactcctc cacagacaga tttgagggga actttgttca cggagaaaag aacggacggg
     1921 ggaagttctt cttctttgat ggcagcaccc tggaggggta ttatgtggat gatgccttgc
     1981 agggccaggg agtttacact tacgaagatg ggggagttct ccagggcacg tatgtagacg
     2041 gagagctgaa cggtccagcc caggaatatg acacagatgg gagactgatc ttcaaggggc
     2101 agtataaaga taacattcgt catggagtgt gctggatata ttacccagat ggaggaagcc
     2161 ttgtaggaga agtaaatgaa gatggggaga tgactggaga gaagatagcc tatgtgtacc
     2221 ctgatgagag gaccgcactt tatgggaaat ttattgatgg agagatgata gaaggcaaac
     2281 tggctaccct tatgtccact gaagaaggga ggcctcactt tgaactgatg cctggaaatt
     2341 cagtgtacca ctttgataag tcgacttcat cttgcatttc taccaatgct cttcttccag
     2401 atccttatga atcagaaagg gtttatgttg ctgaatctct tatttccagt gctggagaag
     2461 gacttttttc aaaggtagct gtgggaccta atactgttat gtctttttat aatggagttc
     2521 gaattacaca ccaagaggtt gacagcaggg actgggccct taatgggaac accctctccc
     2581 ttgatgaaga aacggtcatt gatgtgcctg agccctataa ccacgtatcc aagtactgtg
     2641 cctccttggg acacaaggca aatcactcct tcactccaaa ctgcatcttt gatatgtttg
     2701 tccacccccg ttttgggccc atcaaatgca tccgcaccct gagagcagtg gaggccgatg
     2761 aagagctcac cgttgcctat ggctatgacc acagcccccc cgggaagagt gggcctgaag
     2821 cccctgagtg gtaccaggtg gagctgaagg ccttccaggc cacccagcaa aagtgataat
     2881 actagagcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2941 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     3001 cctttctgcg tttatatact agtagccagt ccggcaaaaa agggcaaggt gtcaccaccc
     3061 tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact
     3121 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3181 atctcgagtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     3241 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3301 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3361 tccataggat ggcaagatcc tggtatcggt ctgtgattcc gactcgtcca acatcaatac
     3421 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3481 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     3541 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3601 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3661 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3721 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3781 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3841 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3901 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3961 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4021 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4081 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     4141 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     4201 aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca
     4261 aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct
     4321 ggctggatga tggggcgatt caggcctggt atgagtcagc aacaccttct tcacgaggca
     4381 gacctcagcg ctagcggagt gtatactggc ttaccatgtt ggcactgatg agggtgtcag
     4441 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     4501 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     4561 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     4621 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     4681 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     4741 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     4801 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     4861 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     4921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     4981 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     5041 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     5101 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     5161 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg
     5221 gtctgacgct cagtggaacg aaaactca
//
