LOCUS       sequence-255035-        8754 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 8754)
  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 8754)
  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: pRS404-PTEF-Ago1
FEATURES             Location/Qualifiers
     source          1..8754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(144..149,150..167,168..174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..198
                     /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     187..209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     206..222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     206..222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     241..261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        243..261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        279..677
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4599..4846
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4615..4633)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4864..4883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4865..4883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4893..4910)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4893..4909)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4902..4924)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5050..5505
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5137..5156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5347..5368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5608..6282)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(6283..6564)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6585..6604)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6704..6726
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6764..6782)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6850..6954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6955..7023,7024..7815)
                     /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(7173..7192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7986..8574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8475..8494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8728..8745
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
       61 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      181 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      241 gcaattaacc ctcactaaag ggaacaaaag ctggagctca tagcttcaaa atgtttctac
      301 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
      361 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
      421 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
      481 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga tttttttctc
      541 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
      601 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
      661 tagcaatcta atctaagttt tctagaacta gtatgtcatc caattcggag gagaacagtc
      721 aagttcctcc ccttgatgcc accgctgctg caactaaacc aaaaaaggcc aagaaaccaa
      781 aggttaagaa gccaaaggat tccgctgagg catcttcatc accagctgct gagggcactg
      841 ctgaagccaa gccaaaaaag gctaagaaat caaagactaa gaagtcaaag gagtctgccg
      901 aggtatcgcc agctccagct gatgaaacta cttctgctgg tgtagatgct aaacctaaga
      961 aggctaaaaa gtcaaaggtc aagaagccaa aggattctac tgaatcatct cctgctccaa
     1021 gtaatgaacc tccagccgcc gaagttgccg ccgaagatgc aaagtctaaa aaggttaaga
     1081 aaccaaaggc caagaaacca aaggaatcta ctgaatcatc tcctgctcca ggtcaggaag
     1141 ccgctgctac tgaaggtgcc acagaagata aacctaagaa ggttaagaaa tcaaaggcta
     1201 agaaggcaaa ggaatctgtt gaatcatctc ctgccgccac tgaatctgtc tctgagaaga
     1261 ctgcaaagaa atctaagaag ccaaaggcta agaaatctac ttctcctgaa acaactgaag
     1321 aaataactga agaatctacc gaatcaaagg aaaagaagac aaagacaaag aaaccaaagg
     1381 aaaagaagtc atctccatct actgccactt cgactgctgc ttcaaaacca gttacttcca
     1441 ttgcgggagt caccattcca ggcaagactt ttgatttgac agatgttcct ccatctaaac
     1501 ctgctccaaa gatggtcccg gaagcctata aattacaaac tagagtggat tacggtacta
     1561 agggtaccaa agtggacgtc ttgactaatc atatactact atctgttggt gatgatgtcc
     1621 ctcaagatga aagagcctct caattggatc catggtggaa atctgcattt gtatatacct
     1681 ataacattac ttttgccgtt ccacagagta attcaccacg taaaggtcca gcgcctgctt
     1741 tatccaaacc aaagaaatac gaattggtag aatctttgtt tcacgaggat gaaaccttgt
     1801 tcaaatataa agatcgtatc tccttcaatg gtgaagatac tctatactct catgtcccat
     1861 tggaggaatt cactttattt gatggttgtt gggatgtcag taacaagcaa aagaaaagaa
     1921 gacaagaagt tgttggtctc aacagtagag ccaaggaaat taatgacttg gctgctcaag
     1981 ttactttaaa atttgctgat aaagtcccat tgggtgatat ttataaggct accacttcaa
     2041 aggatccaga agaacaagaa aataagatgg ctaacgctga taaggttgct ttattgtctt
     2101 tgatgggtgt caaattctta aacacaaagg aacaaatctt ccaacttaac ggtaataaat
     2161 tctttatttt taatgaacac gcaattgcta ctccattcca aattggtggg ttcttaatgc
     2221 atgggttcac agtttcccta cgatatgctt acgggtcagt tttgttaaat actgttaacg
     2281 tctgcctccc atttgttaag tggactaaat atttaccagg tgatgctaaa ttcaaggaaa
     2341 atgaaaagac tcaatacagt ttgttagatt ggattattga atgtatgcac caggccagtg
     2401 ctcaaagagg tcaaaagcta agaggccctc catctgccaa ggatatcaat tttttcattg
     2461 acaagaatag agacattaaa gatctattga agggtttgaa atgttacaga ccatacatta
     2521 attattctgt taacccagat ggaactccaa aaccaccaaa aaagatgcaa gctaagggta
     2581 tcgttggttt cgtaagagag actccagatt ctatgaaatt cagaacttta ccaagtaaca
     2641 tggaaaagaa tggtgtacca aaaccaggtg aaaaggaaat catggtcaca acaaccgcct
     2701 actttgccaa gaagtatgac atcaaattga aatacccaga tgttaaaatg gtgagtctag
     2761 gtggttcgaa tgttgtccca gctgaatgtt taaccattgt tccaggtcag aaattaaagg
     2821 gtttggttta tgacgaaaaa gccgttattg atttcactgc tttaagacca agtgaaaaat
     2881 tcagagccat caccaacttg gcattacctg ccattaagag agctttatca actgaagaag
     2941 aaaatgctaa ggctccacat gattctggtt acactttcat gaaggtccca tctcgtgtca
     3001 ttgacgctcc tgtggttcaa ttcaagaaca ctacagtcac atatgtggat aaaccatttg
     3061 gtactaagaa tggtaagaat aatcatgaag aaactaaggg taattggaat ttaaaggatc
     3121 ataaattcat tactgttcca aaggaaccaa tgcacttgag agctatcttt attaatgatt
     3181 ctgataagtc tccaccagtt tctgtcatgg atgaattgaa ggcctctttg agcaaattcg
     3241 ctgaagatgt cgctgatgtc ggtgttaact ttgacgtatc catggcccca attttaatta
     3301 acaattttaa tgccccaata aagaaggtta ctggtggctt tggtggtaga ggtggacgtg
     3361 gtggtagagg cggccgtggt ggccgtggtg gtagaggagg tcgtggtggt ttcggcggtg
     3421 gccgtggtga aactacttat gaattaactc caggtgaaga aaaactacgt cacttgttag
     3481 ccaacgtccc agaaaagact tatgtcttat tcgtcttggg tcgtggggat gactctgcta
     3541 tttacaacag attgaaatat ttagctgatt tgacgtacgg tgtcattaac aattgtgtta
     3601 tttggaacaa gttcagaaag tgttccactc aatacaatgt taatgtggta atgaagatga
     3661 acttgaagtt agaaggtgct aaccactcct tatgtgcaga agatataaac ttactgaagg
     3721 atgaaaaatc aggattacca ttcatgattt taggtgctga tgttacacat tatccagaaa
     3781 aggatcagaa ttctatctct gcattggtgg gttcttttga tgacaaattt gctcaattcc
     3841 caggttcata tatgttacaa agtggtccag gtgaagaaat aattgctggt atcggtaaca
     3901 tggttctaca gagattgaaa ctgtatcaaa agcacaacaa tggcaaacta cctccaaaga
     3961 ttctattcta cagagatggt gtttctgaat cccaattctc acaaattgtt caaattgaag
     4021 ttaagggttt gaaacaagca ttgaagaaat tcggtagtga attaaacaag ggtgtcaact
     4081 ataacccatc tgtcaccaca atttgtgttg tcaagaggaa tcaaattaga tttatgccac
     4141 ttgagcaaaa tgctataaat gaaaagggtg aagttgctgc cgttcaatca ttcgagaacg
     4201 ttatgccagg tactgttgtt gatcgtggta tcacctctag tgctcacttc gatttcttct
     4261 tgcaatctca acaaccattg aagggtactg gtgttccatg tcattactgg tgtatctacg
     4321 acgaaaatca attcaattct gattacttgc aacaagttac tcacgctttg tgttacttat
     4381 tcggtagatc gagtacaagt attaaggtcg catctcctgt ttattacgct gatttattgt
     4441 gtgaacgtgg tgctgcattc ttcaaggcta actttgagct tgctcaatac gaattttcta
     4501 aggagaggaa gaacagagat gacgttatac cgaccggtaa attactacaa cctgttcata
     4561 agaatgtcac tgacatcatg tactacatat gactcgagtc atgtaattag ttatgtcacg
     4621 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     4681 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     4741 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     4801 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat
     4861 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     4921 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4981 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5041 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5101 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5161 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5221 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5281 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5341 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5401 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5461 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     5521 tctccttacg catctgtgcg gtatttcaca ccgcatagat cggcaagtgc acaaacaata
     5581 cttaaataaa tactactcag taataaccta tttcttagca tttttgacga aatttgctat
     5641 tttgttagag tcttttacac catttgtctc cacacctccg cttacatcaa caccaataac
     5701 gccatttaat ctaagcgcat caccaacatt ttctggcgtc agtccaccag ctaacataaa
     5761 atgtaagctt tcggggctct cttgccttcc aacccagtca gaaatcgagt tccaatccaa
     5821 aagttcacct gtcccacctg cttctgaatc aaacaaggga ataaacgaat gaggtttctg
     5881 tgaagctgca ctgagtagta tgttgcagtc ttttggaaat acgagtcttt taataactgg
     5941 caaaccgagg aactcttggt attcttgcca cgactcatct ccatgcagtt ggacgatatc
     6001 aatgccgtaa tcattgacca gagccaaaac atcctcctta ggttgattac gaaacacgcc
     6061 aaccaagtat ttcggagtgc ctgaactatt tttatatgct tttacaagac ttgaaatttt
     6121 ccttgcaata accgggtcaa ttgttctctt tctattgggc acacatataa tacccagcaa
     6181 gtcagcatcg gaatctagag cacattctgc ggcctctgtg ctctgcaagc cgcaaacttt
     6241 caccaatgga ccagaactac ctgtgaaatt aataacagac atactccaag ctgcctttgt
     6301 gtgcttaatc acgtatactc acgtgctcaa tagtcaccaa tgccctccct cttggccctc
     6361 tccttttctt ttttcgaccg aattaattct taatcggcaa aaaaagaaaa gctccggatc
     6421 aagattgtac gtaaggtgac aagctatttt tcaataaaga atatcttcca ctactgccat
     6481 ctggcgtcat aactgcaaag tacacatata ttacgatgct gttctattaa atgcttccta
     6541 tattatatat atagtaatgt cgttgatcta tggtgcactc tcagtacaat ctgctctgat
     6601 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     6661 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     6721 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     6781 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6841 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6901 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6961 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7021 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7081 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7141 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     7201 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7261 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7321 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7381 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7441 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7501 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7561 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7621 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7681 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7741 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7801 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8101 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     8161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8281 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8641 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8701 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaa
//
