LOCUS       sequence-63936-e        5152 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 5152)
  AUTHORS   Aranko AS, Oeemig JS, Kajander T, Iwai H
  TITLE     Intermolecular domain swapping induces intein-mediated protein 
            alternative splicing.
  JOURNAL   Nat Chem Biol. 2013 Aug 25. doi: 10.1038/nchembio.1320.
  PUBMED    23974115
REFERENCE   2  (bases 1 to 5152)
  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: "Nat Chem 
            Biol. 2013 Aug 25. doi: 10.1038/nchembio.1320."
COMMENT     Sequence Label: pJORSF19
FEATURES             Location/Qualifiers
     source          1..5152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1620..1664
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1702..1720)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1716..1763
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1993..2808)
                     /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(1993..2808)
                     /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     2716..2735
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2809..2900)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2916..3665)
                     /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(2926..2943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3166..3185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3861..4943)
                     /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     4905..4924
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4944..5021)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5067..5086
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5067..5085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5086..5110
                     /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)."
ORIGIN
        1 catggcaatg atcatggtta agaaagggag cgatcctgag gttgttgaga tagatgagat
       61 tgagggactt ggattggaac ttgaagagga gtcaactacc tctaaaaaga aaaagaagga
      121 gaaagagata aagagtattg aggatcttcc aggagtgggt cctgctactg ctgaaaaact
      181 tagggaggcg ggatttgata ctctagaggc tatagcggtt gcatccccta tagaacttaa
      241 agaggttgcg ggaatcagtg aaggggcagc ccttaagata attcaagcgg cgaggaaggc
      301 tgctaatctc gggacgttca tgagagctga cgagtaccta aagaagaggg aaagcattgg
      361 aaggatctca actggaagca aaagtttaga taaattactt ggtggaggta tagagaccca
      421 ggcgataacg gaagtctttg gggagttcgg tagtgggaag tgctttgcta gggataccga
      481 agtttattat gaaaacgata cggtaccaca catggaatca attgaggaga tgtatagtaa
      541 atacgcctcg atgaatgggg aattaccatt cgataatggt tacgcagttc cattagataa
      601 tgtctttgtg tacacgttgg acatcgctag tggtgaaatt aagaaaacga gggcttcata
      661 catctatcgc gagaaggttg agaagctcat tgagataaaa ctatccagtg gatactccct
      721 taaggttact ccctctcacc cagttctcct ctttagagat ggcctgcaat gggtccccgc
      781 tgctgaagtt aaacctggag atgttgttgt tggtgttagg gaagaggtac tgagaagaag
      841 aataatatcc aaaggagaac ttgaattcca tgaggtttcc tcggtaagga taatagatta
      901 caataactgg gtctacgacc ttgtaattcc ggaaacccac aacttcatag ctcccaatgg
      961 acttgttctc cataatactc agctagccca tacccttgcg gtaatggtcc aattaccccc
     1021 agaagaagga ggattaaatg gctcggtaat ttggattgac actgaaaaca ccttcagacc
     1081 tgagaggatt agagagattg cgaagaatag gggattagat cctgacgaag tattaaagca
     1141 catctacgtt gctagggcat ttaatagcaa tcatcaaatg ctgctagttc agcaggccga
     1201 ggataagata aaagagttac tgaacacgga taagcccgtc aagttgctaa tagtggattc
     1261 cctcacgagc cactttagaa gtgaatatat aggtagaggg gcattagctg agagacagca
     1321 gaagttggct aagcatttag ccgatctaca taggttggcg aacctttatg agatagctgt
     1381 ctttgtaacg aaccaagttc aggccaggcc tgatgctttc tttggagatc caacgagacc
     1441 cataggtggg catatcttgg cgcatagtgc aaccttgagg gtctacttaa ggaagggtaa
     1501 gggaggaaag agggtagcta ggttaataga tgcaccacac cttcctgagg gagaggccgt
     1561 tttcaggata acggagaagg gaattgaaga ttaagcttgc ggccgcactc gagtctggta
     1621 aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt
     1681 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1741 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     1801 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     1861 cctgccctga accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga
     1921 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1981 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     2041 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     2101 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2161 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2221 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     2281 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2341 cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca
     2401 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2461 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     2521 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2581 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2641 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     2701 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2761 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt
     2821 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2881 tatttagaaa aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc
     2941 gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg
     3001 gttatccaca gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc
     3061 ggaagaagcc aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3121 aaaatcgacg ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc
     3301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3421 tatcgccact ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc
     3481 acctgttaag gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta
     3541 ccttggttca aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg
     3601 gtttttcttt atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt
     3661 ccgttactct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     3721 atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga
     3781 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     3841 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     3961 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     4021 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4081 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4141 atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4201 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4261 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4321 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4381 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4441 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     4501 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     4561 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     4621 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     4681 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     4741 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     4801 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     4861 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     4921 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     4981 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     5041 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
     5101 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat ac
//
