LOCUS       sequence-55936-e        4542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4542)
  AUTHORS   Ellila S, Jurvansuu JM, Iwai H
  TITLE     Evaluation and comparison of protein splicing by exogenous inteins 
            with foreign exteins in Escherichia coli.
  JOURNAL   FEBS Lett. 2011 Oct 10.
  PUBMED    22001202
REFERENCE   2  (bases 1 to 4542)
  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 Lett. 
            2011 Oct 10."
COMMENT     Sequence Label: pSKDuet26
FEATURES             Location/Qualifiers
     source          1..4542
                     /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             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1096..1140
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1178..1196)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1192..1239
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1469..2284)
                     /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(1469..2284)
                     /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     2192..2211
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2285..2376)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2392..3141)
                     /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(2402..2419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2642..2661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3337..4419)
                     /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     4381..4400
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4420..4497)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagcctagga cagtacaaac ttatcctgaa cggtaaaacc
      181 ctgaaaggtg aaaccaccac cgaagctgtt gacgctgcta ccgcggaaaa agttttcaaa
      241 cagtacgcta acgacaacgg tgttgacggt gaatggacct acgacgacgc taccaaaacc
      301 tacaccgtaa cggaaggatc caagtgcgta tcaggtgaaa caccagttta ccttgcggat
      361 ggcaagacaa taaaaataaa ggatctatac agttctgaga gaaaaaaaga agataacatt
      421 gttgaggctg gttcgggaga agagataata catctaaaag atcccattca aatatattct
      481 tatgtggacg ggaccatagt caggagcaga tcaagacttc tatacaaggg caagagctct
      541 tatcttgtga ggatagaaac tattggcgga agatcggtaa gcgttacacc agttcacaaa
      601 ctctttgtcc ttacggaaaa gggtatcgaa gaggttatgg cctccaacct aaaggtaggc
      661 gacatgattg ctgctgtagc agaaagcgaa tctgaagcaa gagactgcgg aatgagcgag
      721 gaatgcgtga tggaagcaga agtttatacg tcacttgaag cgacattcga tagagtaaag
      781 tctatagcgt acgagaaggg tgattttgat gtatacgatc tttccgtacc agaatacggc
      841 aggaacttta taggcggaga aggacttctc gtacttcaca acacggttat aggtacccag
      901 tacaaactta tcctgaacgg taaaaccctg aaaggtgaaa ccaccaccga agctgttgac
      961 gctgctaccg cggaaaaagt tttcaaacag tacgctaacg acaacggtgt tgacggtgaa
     1021 tggacctacg acgacgctac caaaacctac accgtaacgg aataagcttg gctgttttgg
     1081 cggctcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg
     1141 tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa
     1201 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag
     1261 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     1321 gcggctattt aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag
     1381 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1441 aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact
     1501 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     1561 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     1621 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     1681 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     1741 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     1801 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt
     1861 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     1921 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     1981 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     2041 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     2101 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     2161 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     2221 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     2281 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2341 gatacatatt tgaatgtatt tagaaaaata aacaaatagg catgcagcgc tcttccgctt
     2401 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact
     2461 caaaagcggt aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag
     2521 caaaaagcaa agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc
     2581 cctgacgagc atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta
     2641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2761 tcacgctgtt ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2881 ccggtaagac acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt
     2941 tgtcttgaag ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg
     3001 tcctccaacc cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc
     3061 accgttggta gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag
     3121 tcaacgaaca gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc
     3181 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     3241 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     3301 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3361 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3421 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     3481 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     3541 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     3601 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     3661 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     3721 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     3781 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     3841 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     3901 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     3961 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     4021 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     4081 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     4141 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     4201 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     4261 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     4321 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     4381 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     4441 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     4501 gatctcgacg ctctccctta tgcgactcct gcattaggaa at
//
