LOCUS       sequence-242942-        4289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NpuDnaB-Delta-283-Delta-C39 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4289)
  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 4289)
  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: pHBDuet259
FEATURES             Location/Qualifiers
     source          1..4289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(105..1187)
                     /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     1149..1168
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1188..1265)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1311..1329
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1330..1354
                     /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             1410..1427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1437..1454
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        2001..2019
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2020..2044
                     /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             2153..2197
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2235..2253)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2249..2296
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2526..3341)
                     /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(2526..3341)
                     /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     3249..3268
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3342..3433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3449..4198)
                     /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(3459..3476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3699..3718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
       61 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
      121 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      181 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
      241 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      301 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      361 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
      421 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      481 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      541 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      601 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      661 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      721 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      781 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
      841 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
      901 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
      961 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     1021 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     1081 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     1141 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1201 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1261 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
     1321 cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt tgtttaactt
     1381 taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc agcggcctgg
     1441 tgccgcgcgg cagccatatg cagtacaaac ttatcctgaa cggtaaaacc ctgaaaggtg
     1501 aaaccaccac cgaagctgtt gacgctgcta ccgcggaaaa agttttcaaa cagtacgcta
     1561 acgacaacgg tgttgacggt gaatggacct acgacgacgc taccaaaacc tacaccgtaa
     1621 cggaaggatc cggttgttta gcaggcgata gtctggtgac attagtagat agcggactgc
     1681 aagtgccaat taaggaatta gtaggtaaat ctggttttgc agtttgggca ttgaacgaag
     1741 ctacaatgca gctagaaaag gcaattgtta gcaatgcttt ttcgactggt ataaagcctc
     1801 tgtttacttt gacaactcga ttggggcgga aaatccgtgc cacaggtaat cacaagtttt
     1861 taacaattaa tggctggaaa agacttgatg aattaactcc taaagaacat ctcagtttac
     1921 caagaaatta agcttgcggc cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta
     1981 cacggccgca taatcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     2041 ttccccatct tagtatatta gttaagtata agaaggagat atacatatgg cagatctcaa
     2101 ttggatatcg gccggccacg cgatcgctga cgtcggtacc ctcgagtctg gtaaagaaac
     2161 cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac
     2221 ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     2281 cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc
     2341 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
     2401 tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc
     2461 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     2521 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     2581 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2641 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2701 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2761 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     2821 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2881 gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt acaaacagga
     2941 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3001 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     3061 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3121 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3181 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     3241 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     3301 cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat
     3361 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3421 aaaaataaac aaataggcat gcagcgctct tccgcttcct cgctcactga ctcgctacgc
     3481 tcggtcgttc gactgcggcg agcggtgtca gctcactcaa aagcggtaat acggttatcc
     3541 acagaatcag gggataaagc cggaaagaac atgtgagcaa aaagcaaagc accggaagaa
     3601 gccaacgccg caggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3661 acgctcaagc cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgttggt atctcagttc
     3841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3961 actggcagca gccattggta actgatttag aggactttgt cttgaagtta tgcacctgtt
     4021 aaggctaaac tgaaagaaca gattttggtg agtgcggtcc tccaacccac ttaccttggt
     4081 tcaaagagtt ggtagctcag cgaaccttga gaaaaccacc gttggtagcg gtggtttttc
     4141 tttatttatg agatgatgaa tcaatcggtc tatcaagtca acgaacagct attccgttac
     4201 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     4261 ataagttgta attctcatgt tagtcatgc
//
