LOCUS       sequence-317674-        4179 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 4179)
  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 4179)
  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.119-R5-TEV-SUMO-His
FEATURES             Location/Qualifiers
     source          1..4179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    35..54
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_RNA        57..106
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             230..250
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             284..571
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             572..589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             789..1871
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(808..827)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    1996..2016
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2017..2074
                     /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."
     primer_bind     complement(2079..2096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2250..2838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2330..2349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3009..3800,3801..3869))
                     /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     3632..3651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3870..3974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4042..4060
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4137..4158
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcgctcac aattagctgt
       61 caccggatgt gctttccggt ctgatgagtc cgtgaggacg aaacagcctc tacaaataat
      121 tttgtttaaa gcttacgcgc tctctcttcc tacggttatt ttaatgggta tgagctccaa
      181 aaaatctggt tcatattccg gctctaaagg cagtaaacgt cgtatcttag aaaatctgta
      241 ctttcaaggc tcctccaaaa aatctggttc aggtggtagt ggcgattcag aagtcaatca
      301 agaagctaag ccagaggtca agccagaagt caagcctgag actcacatca atttaaaggt
      361 gtccgatgga tcttcagaga tcttcttcaa gatcaaaaag accactcctt taagaaggct
      421 gatggaagcg ttcgctaaaa gacagggtaa ggaaatggac tccttaagat tcttgtacga
      481 cggtattaga attcaagctg atcaggcccc tgaagatttg gacatggagg ataacgatat
      541 tattgaggct catcgcgaac agattggcgg ccatcatcat caccatcact gataatactg
      601 ctcttcataa ttcagccaaa aaacttaaga ccgccggtct tgtccactac cttgcagtaa
      661 tgcggtggac aggatcggcg gttttctttt ctcttctcaa ccaatggcgg cgcgccatcg
      721 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      781 tggtgaatat gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      841 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
      901 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
      961 gcaaacagtc gttgcttatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     1021 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     1081 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     1141 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     1201 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     1261 gtattatttt ctcccatgag gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     1321 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     1381 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     1441 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     1501 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     1561 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     1621 atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     1681 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     1741 tgttgccagt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     1801 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1861 gcgggcagtg ataattggta acgaatcaga caattgacgg ctcgagggag tagcataggg
     1921 tttgcagaat ccctgcttcg tccatttgac aggcacatta tgcatcgatg ataagctgtc
     1981 aaacatgagc acgcttacta gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc
     2041 accaccctgc cctttttctt taaaaccgaa aagattacct cgctcactga ctcgctgcgc
     2101 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2161 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2221 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2281 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2341 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2401 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2461 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2521 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2581 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2641 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2701 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcggca gattacgcgc
     2821 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2881 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2941 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3001 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3061 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3121 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3181 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3241 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3301 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3361 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3421 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3481 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3541 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3601 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3661 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3721 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3781 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3841 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3901 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3961 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4021 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     4081 ggtgatgacg gtgaaaacct ctgacacatg cagctcttcc ccatcggtga tgtcgggaat
     4141 tcgcggccgc ttctagaggg agaacgatcg ttggctgtg
//
