LOCUS       sequence-36159-e        4581 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 4581)
  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 4581)
  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-KpDcr1delta2d
FEATURES             Location/Qualifiers
     source          1..4581
                     /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(1200..1218)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1214..1261
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1491..2306)
                     /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(1491..2306)
                     /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     2214..2233
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2307..2398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2414..3163)
                     /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(2424..2441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2664..2683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3359..4441)
                     /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     4403..4422
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4442..4519)
                     /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 aaattaaact tcaccaagag attacaaggt aatatcgcta
      961 cacctactag agtggtcaaa gataaaatgt ctaaacgtta tgctgattgt gttgaagcct
     1021 atattggtgc attggttatt gatagatttg gcactgagtt tttggatatc aaggaatggt
     1081 tagaagaact atcagaaaaa aaattggcca aatcgagcca aatgtaactg ctgcgaaatt
     1141 tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac
     1201 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1261 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
     1321 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgacaag
     1381 ctgacgaccg ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     1501 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1561 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1621 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1681 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1741 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1801 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1861 agacgaaata cgcggtcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1921 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1981 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2041 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     2101 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     2161 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     2221 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     2281 gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat
     2341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2401 ggcatgcagc gctcttccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     2461 cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt tatccacaga atcaggggat
     2521 aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc
     2581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagccagag
     2641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2761 aagcgtggcg ctttctcata gctcacgctg ttggtatctc agttcggtgt aggtcgttcg
     2821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccat
     2941 tggtaactga tttagaggac tttgtcttga agttatgcac ctgttaaggc taaactgaaa
     3001 gaacagattt tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa gagttggtag
     3061 ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt ttttctttat ttatgagatg
     3121 atgaatcaat cggtctatca agtcaacgaa cagctattcc gttactctag atttcagtgc
     3181 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
     3241 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     3301 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     3361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3421 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     3481 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     3541 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     3601 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     3661 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     3721 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     3781 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     3841 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     3901 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     3961 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     4021 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     4081 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     4141 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     4201 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     4261 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     4321 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     4381 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     4441 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     4501 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     4561 aaattaatac gactcactat a
//
