LOCUS       sequence-237408-        4398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Charge-Introduced NpuDnaB mini-intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4398)
  AUTHORS   Beyer HM, Mikula KM, Li M, Wlodawer A, Iwai H
  TITLE     The crystal structure of the naturally split gp41-1 intein guides 
            the engineering of orthogonal split inteins from cis-splicing 
            inteins.
  JOURNAL   FEBS J. 2019 Oct 30. doi: 10.1111/febs.15113.
  PUBMED    31665813
REFERENCE   2  (bases 1 to 4398)
  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: "FEBS J. 
            2019 Oct 30. doi: 10.1111/febs.15113."
COMMENT     Sequence Label: pHBDuet139
FEATURES             Location/Qualifiers
     source          1..4398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(124..1206)
                     /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     1168..1187
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1207..1284)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1330..1349
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1330..1348
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1349..1373
                     /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             1429..1446
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1456..1473
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2281..2325
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2363..2381)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2377..2424
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2654..3469)
                     /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(2654..3469)
                     /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     3377..3396
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3470..3561)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3577..4326)
                     /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(3587..3604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3827..3846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
       61 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
      121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
      241 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
      301 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
      361 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
      421 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
      481 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
      541 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
      601 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
      661 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
      721 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
      781 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
      841 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
      901 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
      961 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     1021 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     1081 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     1141 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     1201 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     1261 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     1321 ttaggaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctgtag
     1381 aaataatttt gtttaacttt aataaggaga tataccatgg gcagcagcca tcatcatcat
     1441 catcacagca gcggcctggt gccgcgcggc agcctaggac agtacaaact tatcctgaac
     1501 ggtaaaaccc tgaaaggtga aaccaccacc gaagctgttg acgctgctac cgcggaaaaa
     1561 gttttcaaac agtacgctaa cgacaacggt gttgacggtg aatggaccta cgacgacgct
     1621 accaaaacct acaccgtaac ggaaggatcc ggttgtttag caggcgatag tctggtgaca
     1681 ttagtagata gcggactgca agtgccaatt aaggaattag taggtaaatc tggttttgca
     1741 gtttgggcat tgaacgaagc tacaatgcag ctagaaaagg caattgttag caatgctttt
     1801 tcgactggta aaaagaagct gtttaaattg acaactcgat tggggcggaa aatccgtgcc
     1861 acaggtaatc acaagttttt aacaattaat ggctggaaaa gacttgatga attaactcct
     1921 aaagaacatc tcgctttacc ccggaattct ggtagtgata tttattggga tgaaatagtt
     1981 aaaaaggaat atagtggtga ggaagaagtg tttgacctca ccgttcctgg tttgcataat
     2041 tttgttgcaa ataatattat tgttcacaac tcgggaggta cccagtacaa acttatcctg
     2101 aacggtaaaa ccctgaaagg tgaaaccacc accgaagctg ttgacgctgc taccgcggaa
     2161 aaagttttca aacagtacgc taacgacaac ggtgttgacg gtgaatggac ctacgacgac
     2221 gctaccaaaa cctacaccgt aacggaataa gcttggctgt tttggcggct cgagtctggt
     2281 aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg actcgtctac tagcgcagct
     2341 taattaacct aggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2401 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
     2461 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     2521 ccctgccctg aaccgacgac aagctgacga ccgggtctcc gcaagtggca cttttcgggg
     2581 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2641 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     2701 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     2761 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     2821 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     2881 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     2941 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     3001 tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc gctgttaaaa ggacaattac
     3061 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     3121 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     3181 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     3241 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     3301 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     3361 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     3421 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt
     3481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3541 gtatttagaa aaataaacaa ataggcatgc agcgctcttc cgcttcctcg ctcactgact
     3601 cgctacgctc ggtcgttcga ctgcggcgag cggtgtcagc tcactcaaaa gcggtaatac
     3661 ggttatccac agaatcaggg gataaagccg gaaagaacat gtgagcaaaa agcaaagcac
     3721 cggaagaagc caacgccgca ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3781 aaaaatcgac gctcaagcca gaggtggcga aacccgacag gactataaag ataccaggcg
     3841 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3901 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgttggtat
     3961 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4021 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4081 ttatcgccac tggcagcagc cattggtaac tgatttagag gactttgtct tgaagttatg
     4141 cacctgttaa ggctaaactg aaagaacaga ttttggtgag tgcggtcctc caacccactt
     4201 accttggttc aaagagttgg tagctcagcg aaccttgaga aaaccaccgt tggtagcggt
     4261 ggtttttctt tatttatgag atgatgaatc aatcggtcta tcaagtcaac gaacagctat
     4321 tccgttactc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     4381 catacgatat aagttgta
//
