LOCUS       sequence-100769-        3925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NpuDnaEN11 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3925)
  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 3925)
  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: pSARSF53-756
FEATURES             Location/Qualifiers
     source          1..3925
                     /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             479..523
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(561..579)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      575..622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(852..1667)
                     /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(852..1667)
                     /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     1575..1594
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1668..1759)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1775..2524)
                     /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(1785..1802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2025..2044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2720..3802)
                     /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     3764..3783
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3803..3880)
                     /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 taagcttgcg gccgcactcg agtctggtaa
      481 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
      541 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      601 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
      661 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
      721 ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact tttcggggaa
      781 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      841 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
      901 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
      961 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1021 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1081 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1141 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1201 attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg acaattacaa
     1261 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     1321 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     1381 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     1441 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     1501 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     1561 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     1621 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt
     1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1741 atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct cactgactcg
     1801 ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg
     1861 ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag caaagcaccg
     1921 gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1981 aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gttggtatct
     2161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2281 atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg aagttatgca
     2341 cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca acccacttac
     2401 cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg
     2461 tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga acagctattc
     2521 cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     2581 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     2641 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     2701 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     2821 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     2881 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     2941 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     3001 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     3061 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     3121 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     3181 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     3241 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     3301 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     3361 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     3421 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     3481 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     3541 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     3601 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     3661 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     3721 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     3781 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     3841 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     3901 ttatgcgact cctgcattag gaaat
//
