LOCUS       sequence-256079-        7897 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 7897)
  AUTHORS   Drinnenberg IA, Weinberg DE, Xie KT, Mower JP, Wolfe KH, Fink GR, 
            Bartel DP
  TITLE     RNAi in budding yeast.
  JOURNAL   Science. 2009 Oct 23;326(5952):544-550. doi: 
            10.1126/science.1176945. Epub 2009 Sep 10.
  PUBMED    19745116
REFERENCE   2  (bases 1 to 7897)
  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"; doi: 
            "10.1126/science.1176945"; journalName: "Science"; date: 
            "2009-10-23- 23"; volume: "326"; issue: "5952"; pages: "544-550"
COMMENT     Sequence Label: pRS405-PTEF-Dcr1
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..88
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(103..108,109..126,127..133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..157
                     /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)."
     primer_bind     146..168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     165..181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     165..181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     200..220
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        202..220
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        238..636
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      2484..2731
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2500..2518)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2749..2768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2750..2768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2778..2795)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2778..2794)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2787..2809)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2935..3390
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3022..3041)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3232..3253
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3691..4098
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4099..5193
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(5687..5706)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5806..5828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5866..5884)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5952..6056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6057..6125,6126..6917)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6275..6294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7088..7676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7577..7596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7830..7847
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
       61 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      121 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      181 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctcat
      241 agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc
      301 gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta
      361 gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc
      421 tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt
      481 tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc
      541 ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt
      601 gctcattaga aagaaagcat agcaatctaa tctaagtttt ctagatgaat agagaaaaaa
      661 gcgccgatct aagtaaaaca tcagatgctc catacaatga actcgatgcc aaaaatttaa
      721 gaaatttcta taaggttcaa aatgcatgtg cgcaattgag ggaatccata aaagtaattt
      781 acgaaaacgg gttatcatcc gatcaattga atactatggc gaaacatggg aacgacctag
      841 aaaaatctat tgcaaacagt cctgctatgt ccgtagcaag ttgcttgaac caagttagac
      901 ccaccctaga catcaagaat atctttgatc attataaatt tgaaaataat tcatcaccag
      961 ttgatcctta tgtccattat cccgtatgtt ctgatcaaaa tttggaaaat ttggctttca
     1021 ttcatagatc tttgccgaat atgaatgtca aattgactga attgcaaaag accgtaatga
     1081 gtaatgaacg tttggaattt ttaggtgata gttggcttgg cgccttagtg gcgtatatta
     1141 tttataagaa atatccttat gctaatgaag gtgccttatc aaagatgaag gaagctatcg
     1201 ttaacaacaa caatttggaa aagatctgtg aaaaacttgg ctttaaagaa agattaaagg
     1261 aaaatattcc acgttcttca atgaaaatca aagatagatt aaccaagaat tacgcagact
     1321 gtgtggaagc ctacattggc gcattggtta ttgatagatt ttcaacagaa tttaatgatg
     1381 ttgctctttg gcttgaagaa ctatctgagg aacatttcat tgaactagga cctatgatgg
     1441 tcaaagagcc tttaaacaaa aatgcaaagg gtgaactagg tgcgttctta caatttaata
     1501 atattggagc caaaatatca tataaaagat tgaatgataa atctccgttt aaggtagaag
     1561 taagattggg gaataatttg ttaggtattg gagatggttc aaatgtgaga gaagctgaac
     1621 agagagccgc tatggaagca cttgctcaac cgaaattgat tcaaaaatat tctctacatg
     1681 atattgaact agaaagaggc atgattgaag aagcagataa tgttccccaa ctactcaaaa
     1741 gtgcggaaat tcccaatcaa acaccacaaa gtcatcattc tatcacagaa gaaattgatg
     1801 aagaagaagg tcttacattc ccccattcga atccattttt gaaaggtaca ccaccagcag
     1861 ttttacctga aattgtaact aatcaaattc cacaggcttc tgcggatcat caagaacaaa
     1921 atgtgcctga cactgataaa attgtgaatg atgttatgga acgtatgagt aagatactgt
     1981 cggtgatggt ttctgaggct gtttcaaatg cacttggtag accagctcca aaaaatgcac
     2041 ctatccctat cccagtaaca accccagtct caccgctagc aacatcccca gtctcagtct
     2101 cagtcccagc gccagctcca gccgtagctg tcaccccatc agttgtgaat aatccaccac
     2161 aacccaaaat ggtcacacaa ccggtaagca caaacctaaa taaaacgcct gaagttgcac
     2221 ttttaaattc tgtgtatgat aaagaagcct cagggagact ttatgcattg ttaggtaaat
     2281 ataaattgta tccagaatat aataccgagc aactaggaat gacagatttt tatacggtat
     2341 gctttataaa gggtgttggt gtagaaattg gtaaaggcca tggtagaagt aaaaagatat
     2401 cccaacataa atctgcagaa gatgctttga atggaaaagc actgaaggaa tacttgaggc
     2461 attgcaacaa tctgtgactc gagtcatgta attagttatg tcacgcttac attcacgccc
     2521 tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     2581 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     2641 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     2701 ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc ctatagtgag
     2761 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2821 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2881 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     2941 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3001 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3061 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3121 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3181 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3241 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3301 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3361 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     3421 gtgcggtatt tcacaccgca tatcgaccgg tcgaggagaa cttctagtat atctacatac
     3481 ctaatattat tgccttatta aaaatggaat cccaacaatt acatcaaaat ccacattctc
     3541 ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt tcaaaaacgt tatatttata
     3601 ggataattat actctatttc tcaacaagta attggttgtt tggccgagcg gtctaaggcg
     3661 cctgattcaa gaaatatctt gaccgcagtt aactgtggga atactcaggt atcgtaagat
     3721 gcaagagttc gaatctctta gcaaccatta tttttttcct caacataacg agaacacaca
     3781 ggggcgctat cgcacagaat caaattcgat gactggaaat tttttgttaa tttcagaggt
     3841 cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa ggtgagagcg
     3901 ccggaaccgg cttttcatat agaatagaga agcgttcatg actaaatgct tgcatcacaa
     3961 tacttgaagt tgacaatatt atttaaggac ctattgtttt ttccaatagg tggttagcaa
     4021 tcgtcttact ttctaacttt tcttaccttt tacatttcag caatatatat atatatattt
     4081 caaggatata ccattctaat gtctgcccct aagaagatcg tcgttttgcc aggtgaccac
     4141 gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc tgatgttcgt
     4201 tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat cgatgctaca
     4261 ggtgttccac ttccagatga ggcgctggaa gcctccaaga aggctgatgc cgttttgtta
     4321 ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta gacctgaaca aggtttacta
     4381 aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt tgcatccgac
     4441 tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga cttcgttgtt
     4501 gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagacga tggtgatggt
     4561 gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac aagaatggcc
     4621 gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga taaagctaat
     4681 gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa gaacgaattc
     4741 cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct agttaagaac
     4801 ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat catctccgat
     4861 gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg
     4921 ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc tccagatttg
     4981 ccaaagaata aggtcaaccc tatcgccact atcttgtctg ctgcaatgat gttgaaattg
     5041 tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa ggttttggat
     5101 gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt cggtgatgct
     5161 gtcgccgaag aagttaagaa aatccttgct taaaaagatt ctcttttttt atgatatttg
     5221 tacataaact ttataaatga aattcataat agaaacgaca cgaaattaca aaatggaata
     5281 tgttcatagg gtagacgaaa ctatatacgc aatctacata catttatcaa gaaggagaaa
     5341 aaggaggatg taaaggaata caggtaagca aattgatact aatggctcaa cgtgataagg
     5401 aaaaagaatt gcactttaac attaatattg acaaggagga gggcaccaca caaaaagtta
     5461 ggtgtaacag aaaatcatga aactatgatt cctaatttat atattggagg attttctcta
     5521 aaaaaaaaaa aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta
     5581 agtcaatacc ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc
     5641 tgtcagaaac ggccttaacg acgtagtcga tatggtgcac tctcagtaca atctgctctg
     5701 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5761 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5821 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5881 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5941 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6001 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6061 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6121 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6181 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6241 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6301 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6361 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6421 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6481 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6541 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6601 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6661 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6721 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6781 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6841 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6901 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6961 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7021 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7081 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7141 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7201 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7261 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7321 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7381 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7441 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7501 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7561 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7621 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7681 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7741 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7801 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7861 gcccaatacg caaaccgcct ctccccgcgc gttggcc
//
