LOCUS       sequence-36183-e        4857 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 4857)
  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 4857)
  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-2)
FEATURES             Location/Qualifiers
     source          1..4857
                     /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(1476..1494)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1490..1537
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1767..2582)
                     /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(1767..2582)
                     /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     2490..2509
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2583..2674)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2690..3439)
                     /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(2700..2717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2940..2959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3635..4717)
                     /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     4679..4698
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4718..4795)
                     /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 cataaaagtt
      721 ttccaaatat gaatgcacat ctaaatgatg ctcaaaagac tcaattatct aatgaaaggt
      781 tagaattttt gggtgatagt tggctaggtg ctttagtttc atatattgta tacaccagat
      841 ttccttctgc aaatgaaggt atgctttctc aaatgaaaga atcaatagta aataataata
      901 atctatttga ttggagtaca aaattacctt ctgtagtttc tgaggttatt atagaatccc
      961 atccgaaaat acagcctgac tctaaacgtt atgctgattg tgttcaagcc tatattggtg
     1021 cattggttat tgatagattt ggcactgagt ttttggatat caaggaatgg ttagaagaac
     1081 tatcagaaaa aaaattggcc aaatcgagcc aaatggtgat aaaggaacca ttaaataaaa
     1141 atgctaaaaa tgaattagcc gaattactgc aaattaacaa attaggacat aaacttcatt
     1201 acagaaagct tacagaaatg cctccattca gagttgaagt taaaataggt gacattctgt
     1261 tagacgaagc tgaaggtaac agtataaggg aagcagaaca cagagctgcg atgaaggtat
     1321 tggaaaatga tgaattattg aggaaatatt cggtttatga tttagaggat ggaaactcga
     1381 agaaatattc tattgaagag taactgctgc gaaatttgaa cgccagcaca tggactcgtc
     1441 tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
     1501 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
     1561 gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
     1621 ggctatttaa cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg
     1681 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1741 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     1801 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     1861 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     1921 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     1981 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     2041 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     2101 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta
     2161 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     2221 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     2281 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     2341 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     2401 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     2461 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2521 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     2581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2641 tacatatttg aatgtattta gaaaaataaa caaataggca tgcagcgctc ttccgcttcc
     2701 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggtgtc agctcactca
     2761 aaagcggtaa tacggttatc cacagaatca ggggataaag ccggaaagaa catgtgagca
     2821 aaaagcaaag caccggaaga agccaacgcc gcaggcgttt ttccataggc tccgcccccc
     2881 tgacgagcat cacaaaaatc gacgctcaag ccagaggtgg cgaaacccga caggactata
     2941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3061 acgctgttgg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3181 ggtaagacac gacttatcgc cactggcagc agccattggt aactgattta gaggactttg
     3241 tcttgaagtt atgcacctgt taaggctaaa ctgaaagaac agattttggt gagtgcggtc
     3301 ctccaaccca cttaccttgg ttcaaagagt tggtagctca gcgaaccttg agaaaaccac
     3361 cgttggtagc ggtggttttt ctttatttat gagatgatga atcaatcggt ctatcaagtc
     3421 aacgaacagc tattccgtta ctctagattt cagtgcaatt tatctcttca aatgtagcac
     3481 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     3541 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     3601 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3721 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3781 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3841 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3901 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3961 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4021 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4081 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4141 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4201 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4261 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4321 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4381 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4441 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4501 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4561 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4621 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4681 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4741 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4801 tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact cactata
//
