LOCUS       sequence-36179-e        4815 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 4815)
  AUTHORS   Weinberg DE, Nakanishi K, Patel DJ, Bartel DP
  TITLE     The inside-out mechanism of dicers from budding yeasts.
  JOURNAL   Cell. 2011 Jul 22;146(2):262-76.
  PUBMED    21784247
REFERENCE   2  (bases 1 to 4815)
  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: "Cell."; 
            date: "2011-07-22"; volume: "146(2)"; pages: "262-76"
COMMENT     Sequence Label: pRSF-KpDcr1deltaC(E224Q, GiVL-1)
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..50
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             60..77
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             90..383
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             505..516
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1434..1452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1448..1495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1725..2540)
                     /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(1725..2540)
                     /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     2448..2467
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2541..2632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2648..3397)
                     /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(2658..2675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2898..2917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3593..4675)
                     /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     4637..4656
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4676..4753)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctttaataag gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc
       61 tggtgccgcg cggcagccat atggctagca tgtcggactc agaagtcaat caagaagcta
      121 agccagaggt caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg
      181 gatcttcaga gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag
      241 cgttcgctaa aagacagggt aaggaaatgg actccttaag attcttgtac gacggtatta
      301 gaatccaagc tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg
      361 ctcacagaga acagattggt ggatccagca acgagttaaa agttagagaa ttttatagac
      421 tccacaatgc ttgtgtgaaa ttgaaagagt ctattaaatt gatttatgaa aacccattgg
      481 taactgacca aaatgttttg aatttgggaa ctgcagaaaa tactattgat tatacaattt
      541 taaacacacc tacactaaat gttgctaaaa ctttactggg caatagatat tcattagatc
      601 taatagatct ttttcaatct catgatttca aagattcaaa cactgatgtg gatatgttta
      661 ttaaatatcc tgtggtttat gatgaaaatt tggaaaattt agcatttatg cctcacgtgt
      721 acgaaaggtt agaatttttg ggtgatagtt ggctaggtgc tttagtttca tatattgtat
      781 acaccagatt tccttctgca aatgaaggta tgctttctca aatgaaagaa tcaatagtaa
      841 ataataataa tctatttgat tggagtacaa aattaaactt caccaagaga ttacaaggta
      901 atatcgctac acctactaga gtggtcaaag ataaaatgtc taaacgttat gctgattgtg
      961 ttgaagccta tattggtgca ttggttattg atagatttgg cactgagttt ttggatatca
     1021 aggaatggtt agaagaacta tcagaaaaaa aattggccaa atcgagccaa atggtgataa
     1081 aggaaccatt aaataaaaat gctaaaaatg aattagccga attactgcaa attaacaaat
     1141 taggacataa acttcattac agaaagctta cagaaatgcc tccattcaga gttgaagtta
     1201 aaataggtga cattctgtta gacgaagctg aaggtaacag tataagggaa gcagaacaca
     1261 gagctgcgat gaaggtattg gaaaatgatg aattattgag gaaatattcg gtttatgatt
     1321 tagaggatgg aaactcgaag aaatattcta ttgaagagta actgctgcga aatttgaacg
     1381 ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg ccaccgctga
     1441 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1501 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1561 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga caagctgacg
     1621 accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1681 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     1741 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     1801 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     1861 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1921 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1981 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     2041 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     2101 aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     2161 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     2221 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     2281 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     2341 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     2401 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     2461 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     2521 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggcatg
     2641 cagcgctctt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     2701 gcggtgtcag ctcactcaaa agcggtaata cggttatcca cagaatcagg ggataaagcc
     2761 ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag ccaacgccgc aggcgttttt
     2821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagcc agaggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3001 ggcgctttct catagctcac gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa
     3061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccattggtaa
     3181 ctgatttaga ggactttgtc ttgaagttat gcacctgtta aggctaaact gaaagaacag
     3241 attttggtga gtgcggtcct ccaacccact taccttggtt caaagagttg gtagctcagc
     3301 gaaccttgag aaaaccaccg ttggtagcgg tggtttttct ttatttatga gatgatgaat
     3361 caatcggtct atcaagtcaa cgaacagcta ttccgttact ctagatttca gtgcaattta
     3421 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     3481 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3541 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     3601 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3661 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     3721 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3781 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     3841 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     3901 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     3961 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4021 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     4081 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     4141 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     4201 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     4261 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     4321 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     4381 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     4441 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     4501 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     4561 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4621 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     4681 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4741 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta
     4801 atacgactca ctata
//
