LOCUS       sequence-100773-        3934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NpuDnaB(delta283)N11 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3934)
  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 3934)
  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-HY100
FEATURES             Location/Qualifiers
     source          1..3934
                     /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             488..532
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(570..588)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      584..631
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(861..1676)
                     /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(861..1676)
                     /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     1584..1603
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1677..1768)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1784..2533)
                     /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(1794..1811)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2034..2053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2729..3811)
                     /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     3773..3792
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3812..3889)
                     /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 tggatccggt
      421 tgtttagcag gcgatagtct ggtgacatta gtagatagct aagcttgcgg ccgcactcga
      481 gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag
      541 cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      601 ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac
      661 ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat
      721 ttaacgaccc tgccctgaac cgacgacaag ctgacgaccg ggtctccgca agtggcactt
      781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      841 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      901 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      961 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1021 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1081 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     1141 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1201 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct gttaaaagga
     1261 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1321 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     1381 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1441 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1501 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     1561 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1621 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc
     1681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1741 tttgaatgta tttagaaaaa taaacaaata ggcatgcagc gctcttccgc ttcctcgctc
     1801 actgactcgc tacgctcggt cgttcgactg cggcgagcgg tgtcagctca ctcaaaagcg
     1861 gtaatacggt tatccacaga atcaggggat aaagccggaa agaacatgtg agcaaaaagc
     1921 aaagcaccgg aagaagccaa cgccgcaggc gtttttccat aggctccgcc cccctgacga
     1981 gcatcacaaa aatcgacgct caagccagag gtggcgaaac ccgacaggac tataaagata
     2041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2161 ttggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2281 acacgactta tcgccactgg cagcagccat tggtaactga tttagaggac tttgtcttga
     2341 agttatgcac ctgttaaggc taaactgaaa gaacagattt tggtgagtgc ggtcctccaa
     2401 cccacttacc ttggttcaaa gagttggtag ctcagcgaac cttgagaaaa ccaccgttgg
     2461 tagcggtggt ttttctttat ttatgagatg atgaatcaat cggtctatca agtcaacgaa
     2521 cagctattcc gttactctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     2581 tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa
     2641 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     2701 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2761 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2821 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     2881 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     2941 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     3001 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     3061 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     3121 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     3181 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     3241 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     3301 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     3361 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     3421 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     3481 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     3541 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     3601 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     3661 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     3721 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     3781 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     3841 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     3901 cgctctccct tatgcgactc ctgcattagg aaat
//
