LOCUS       sequence-100767-        3997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NpuDnaEN35 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3997)
  AUTHORS   Aranko AS, Oeemig JS, Zhou D, Kajander T, Wlodawer A, Iwai H
  TITLE     Structure-based engineering and comparison of novel split inteins 
            for protein ligation.
  JOURNAL   Mol Biosyst. 2014 May;10(5):1023-34. doi: 10.1039/c4mb00021h.
  PUBMED    24574026
REFERENCE   2  (bases 1 to 3997)
  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"; doi: "10"; journalName: 
            "Mol Biosyst"; date: "2014-05"; volume: "10"; issue: "5"; pages: 
            "1023-34"
COMMENT     Sequence Label: pSARSF53-218
FEATURES             Location/Qualifiers
     source          1..3997
                     /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"
     protein_bind    20..44
                     /label=lac operator
                     /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)."
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             124..414
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIETHREQIGG"
     CDS             551..595
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(633..651)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      647..694
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(924..1739)
                     /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(924..1739)
                     /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     1647..1666
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1740..1831)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1847..2596)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(1857..1874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2097..2116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2792..3874)
                     /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     3836..3855
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3875..3952)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 actagtgact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
      181 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag aaatcttctt taagatcaaa
      241 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
      301 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
      361 ttggacatgg aggataacga tattattgag actcacagag aacagattgg tggatcctgt
      421 ttaagctatg aaacggaaat attgacagta gaatatggat tattaccgat tggtaaaatt
      481 gtagaaaagc gcatcgaatg tactgtttat agcgttgata attaagcttg cggccgcact
      541 cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg actcgtctac
      601 tagcgcagct taattaacct aggctgctgc caccgctgag caataactag cataacccct
      661 tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca
      721 cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc
      781 tatttaacga ccctgccctg aaccgacgac aagctgacga ccgggtctcc gcaagtggca
      841 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      901 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
      961 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     1021 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     1081 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     1141 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     1201 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     1261 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc gctgttaaaa
     1321 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     1381 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     1441 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     1501 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     1561 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     1621 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     1681 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     1741 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1801 atatttgaat gtatttagaa aaataaacaa ataggcatgc agcgctcttc cgcttcctcg
     1861 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggtgtcagc tcactcaaaa
     1921 gcggtaatac ggttatccac agaatcaggg gataaagccg gaaagaacat gtgagcaaaa
     1981 agcaaagcac cggaagaagc caacgccgca ggcgtttttc cataggctcc gcccccctga
     2041 cgagcatcac aaaaatcgac gctcaagcca gaggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2221 ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2341 aagacacgac ttatcgccac tggcagcagc cattggtaac tgatttagag gactttgtct
     2401 tgaagttatg cacctgttaa ggctaaactg aaagaacaga ttttggtgag tgcggtcctc
     2461 caacccactt accttggttc aaagagttgg tagctcagcg aaccttgaga aaaccaccgt
     2521 tggtagcggt ggtttttctt tatttatgag atgatgaatc aatcggtcta tcaagtcaac
     2581 gaacagctat tccgttactc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     2641 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
     2701 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     2761 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2881 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     2941 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     3001 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     3061 ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     3121 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     3181 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     3241 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     3301 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     3361 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     3421 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     3481 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     3541 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     3601 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     3661 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     3721 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     3781 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     3841 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     3901 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     3961 cgacgctctc ccttatgcga ctcctgcatt aggaaat
//
