LOCUS       sequence-242939-        4289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Charge-introduced 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: pSADuet148
FEATURES             Location/Qualifiers
     source          1..4289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..48
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    49..73
                     /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             129..146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             156..173
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        720..738
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    739..763
                     /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             872..916
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(954..972)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      968..1015
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1245..2060)
                     /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(1245..2060)
                     /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     1968..1987
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2061..2152)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2168..2917)
                     /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(2178..2195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2418..2437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3113..4195)
                     /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     4157..4176
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4196..4273)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tcccttatgc gactcctgca ttaggaaatt aatacgactc actatagggg aattgtgagc
       61 ggataacaat tcccctgtag aaataatttt gtttaacttt aataaggaga tataccatgg
      121 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgc
      181 agtacaaact tatcctgaac ggtaaaaccc tgaaaggtga aaccaccacc gaagctgttg
      241 acgctgctac cgcggaaaaa gttttcaaac agtacgctaa cgacaacggt gttgacggtg
      301 aatggaccta cgacgacgct accaaaacct acaccgtaac ggaaggatcc ggttgtttag
      361 caggcgatag tctggtgaca ttagtagata gcggactgca agtgccaatt aaggaattag
      421 taggtaaatc tggttttgca gtttgggcat tgaacgaagc tacaatgcag ctagaaaagg
      481 caattgttag caatgctttt tcgactggta aaaagaagct gtttaaattg acaactcgat
      541 tggggcggaa aatccgtgcc acaggtaatc acaagttttt aacaattaat ggctggaaaa
      601 gacttgatga attaactcct aaagaacatc tcagtttacc aagaaattaa gcttgcggcc
      661 gcataatgct taagtcgaac agaaagtaat cgtattgtac acggccgcat aatcgaaatt
      721 aatacgactc actatagggg aattgtgagc ggataacaat tccccatctt agtatattag
      781 ttaagtataa gaaggagata tacatatggc agatctcaat tggatatcgg ccggccacgc
      841 gatcgctgac gtcggtaccc tcgagtctgg taaagaaacc gctgctgcga aatttgaacg
      901 ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg ccaccgctga
      961 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1021 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1081 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga caagctgacg
     1141 accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1201 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     1261 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     1321 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     1381 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1441 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1501 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1561 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1621 aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1681 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1741 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1801 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1861 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1921 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1981 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     2041 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggcatg
     2161 cagcgctctt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     2221 gcggtgtcag ctcactcaaa agcggtaata cggttatcca cagaatcagg ggataaagcc
     2281 ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag ccaacgccgc aggcgttttt
     2341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagcc agaggtggcg
     2401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2521 ggcgctttct catagctcac gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa
     2581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccattggtaa
     2701 ctgatttaga ggactttgtc ttgaagttat gcacctgtta aggctaaact gaaagaacag
     2761 attttggtga gtgcggtcct ccaacccact taccttggtt caaagagttg gtagctcagc
     2821 gaaccttgag aaaaccaccg ttggtagcgg tggtttttct ttatttatga gatgatgaat
     2881 caatcggtct atcaagtcaa cgaacagcta ttccgttact ctagatttca gtgcaattta
     2941 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     3001 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3061 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     3121 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3181 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     3241 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3301 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     3361 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     3421 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     3481 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     3541 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     3601 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     3661 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     3721 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     3781 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     3841 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     3901 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     3961 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     4021 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     4081 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4141 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     4201 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4261 attcgatggt gtccgggatc tcgacgctc
//
