LOCUS       sequence-9509-ex        4726 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 4726)
  AUTHORS   Muona M, Aranko AS, Iwai H
  TITLE     Segmental isotopic labelling of a multidomain protein by protein 
            ligation by protein trans-splicing.
  JOURNAL   Chembiochem. 2008 Dec 15. 9(18):2958-61.
  PUBMED    19031436
REFERENCE   2  (bases 1 to 4726)
  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: 
            "Chembiochem. 2008 Dec 15. 9(18):2958-61."
COMMENT     Sequence Label: pMMRSF17
FEATURES             Location/Qualifiers
     source          1..4726
                     /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             124..414
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1280..1324
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1362..1380)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1376..1423
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1653..2468)
                     /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(1653..2468)
                     /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     2376..2395
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2469..2560)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2576..3325)
                     /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(2586..2603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2826..2845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3521..4603)
                     /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     4565..4584
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4604..4681)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 actagtgact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
      181 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag aaatcttctt taagatcaaa
      241 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
      301 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
      361 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggttcgaag
      421 agccatatgc aggaggaggc agagtatgtg cgggccctct ttgactttaa tgggaatgat
      481 gaagaagatc ttccctttaa gaaaggagac atcctgagaa tccgggataa gcctgaagag
      541 cagtggtgga atgcagagga cagcgaagga aagaggggga tgattcctgt cccttacgtg
      601 gagaagtata gacctgcctc cgcctcagta tcggctctga ttggaggtaa ccagggatcc
      661 tgtttaagct atgaaacgga aatattgaca gtagaatatg gattattacc gattggtaaa
      721 attgtagaaa agcgcatcga atgtactgtt tatagcgttg ataataatgg aaatatttat
      781 acacaacctg tagcacaatg gcacgatcgc ggagaacaag aggtgtttga gtattgtttg
      841 gaagatggtt cattgattcg ggcaacaaaa gaccataagt ttatgactgt tgatggtcaa
      901 atgttgccaa ttgatgaaat atttgaacgt gaattggatt tgatgcgggt tgataatttg
      961 ccgaatatca aaatagccac acgtaaatat ttaggcaaac aaaatgtcta tgacattgga
     1021 gttgagcgct aacataattt tgcactcaaa aatggcttca tagcttctaa ttgtttcaat
     1081 ggtacccagt acaaacttat cctgaacggt aaaaccctga aaggtgaaac caccaccgaa
     1141 gctgttgacg ctgctaccgc ggaaaaagtt ttcaaacagt acgctaacga caacggtgtt
     1201 gacggtgaat ggacctacga cgacgctacc aaaacctaca ccgtaacgga ataagcttgc
     1261 ggccgcactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga
     1321 ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc aataactagc
     1381 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt
     1441 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
     1501 ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac cgggtctccg
     1561 caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1621 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
     1681 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     1741 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     1801 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     1861 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     1921 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     1981 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg
     2041 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2101 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     2161 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     2221 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     2281 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     2341 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     2401 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     2461 tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca gcgctcttcc
     2581 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct
     2641 cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg aaagaacatg
     2701 tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc ataggctccg
     2761 cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa acccgacagg
     2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2941 tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3061 caacccggta agacacgact tatcgccact ggcagcagcc attggtaact gatttagagg
     3121 actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat tttggtgagt
     3181 gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga accttgagaa
     3241 aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca atcggtctat
     3301 caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc tcttcaaatg
     3361 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     3421 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3481 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3601 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3661 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3721 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     3781 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     3841 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     3901 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     3961 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4021 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4081 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4141 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4201 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4261 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4321 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4381 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4441 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4501 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4561 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4621 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4681 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaat
//
