LOCUS       sequence-66338-e        4696 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 4696)
  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 4696)
  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: pSARSF505
FEATURES             Location/Qualifiers
     source          1..4696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     319..338
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(358..435)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     481..500
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        481..499
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    500..524
                     /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             580..597
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(631..659)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1665..1685
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1730..1774
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1812..1830)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1826..1873
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2103..2918)
                     /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(2103..2918)
                     /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     2826..2845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2919..3010)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3026..3775)
                     /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(3036..3053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3276..3295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3971..4696,1..357))
                     /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"
ORIGIN
        1 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
       61 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
      121 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
      181 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
      241 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
      301 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
      361 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
      421 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat
      481 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
      541 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      601 actagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      661 gatgttaatg ggcacaaatt ttctgtccgc ggagagggtg aaggtgatgc tacaaacgga
      721 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacacta
      781 gttactactt tatgtttaag ctatgaaacg gaaatattga cagtagaata tggattatta
      841 ccgattggta aaattgtaga aaagcgcatc gaatgtactg tttatagcgt tgataataat
      901 ggaaatattt atacacaacc tgtagcacaa tggcacgatc gcggagaaca agaggtgttt
      961 gagtattgtt tggaagatgg ttcattgatt cgggcaacaa aagaccataa gtttatgact
     1021 gttgatggtc aaatgttgcc aattgatgaa atatttgaac gtgaattgga tttgatgcgg
     1081 gttgataatt tgccgaatat caaaatagcc acacgtaaat atttaggcaa acaaaatgtc
     1141 tatgacattg gagttgagcg cgaccataat tttgcactca aaaatggctt catagcttct
     1201 aattgttatg gtgttcaatg cttttctaga tatccggatc atatgaaacg gcatgacttt
     1261 ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt caaagatgac
     1321 gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt taatcgtatc
     1381 gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa actcgagtac
     1441 aactataact cacacaatgt atacatcacg gcagacaaac aaaagaatgg aatcaaagct
     1501 aacttcaaaa ttcgccacaa cattgaagat ggatccgttc aactagcaga ccattatcaa
     1561 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtcgaca
     1621 caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct tcttgagttt
     1681 gtaactgctg ctgggttgac ttaagcttgc ggccgcactc gagtctggta aagaaaccgc
     1741 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     1801 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1861 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     1921 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     1981 accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga aatgtgcgcg
     2041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     2101 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2161 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2221 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2281 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2341 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2401 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2461 tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca aacaggaatc
     2521 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2581 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     2641 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2701 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2761 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2821 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2881 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     2941 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3001 aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc gctacgctcg
     3061 gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg gttatccaca
     3121 gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc
     3181 aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3241 ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc tcagttcggt
     3421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3541 ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc acctgttaag
     3601 gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta ccttggttca
     3661 aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg gtttttcttt
     3721 atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt ccgttactct
     3781 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     3841 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3901 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3961 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4021 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     4081 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4141 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     4201 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     4261 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     4321 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     4381 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     4441 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     4501 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     4561 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4621 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4681 taatgatcag cccact
//
