LOCUS       sequence-10224-e        8745 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 8745)
  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 8745)
  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..8745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2241..2261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2279..2677
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      6599..6846
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6615..6633)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6864..6883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6865..6883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6893..6910)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6893..6909)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6902..6924)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7050..7505
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7137..7156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7347..7368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7604..8278)
                     /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(8279..8559)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8576..8595)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8695..8717
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctca
     2281 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     2341 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     2401 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     2461 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     2521 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
     2581 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
     2641 tgctcattag aaagaaagca tagcaatcta atctaagttt tctagaacta gtatgtcatc
     2701 caattcggag gagaacagtc aagttcctcc ccttgatgcc accgctgctg caactaaacc
     2761 aaaaaaggcc aagaaaccaa aggttaagaa gccaaaggat tccgctgagg catcttcatc
     2821 accagctgct gagggcactg ctgaagccaa gccaaaaaag gctaagaaat caaagactaa
     2881 gaagtcaaag gagtctgccg aggtatcgcc agctccagct gatgaaacta cttctgctgg
     2941 tgtagatgct aaacctaaga aggctaaaaa gtcaaaggtc aagaagccaa aggattctac
     3001 tgaatcatct cctgctccaa gtaatgaacc tccagccgcc gaagttgccg ccgaagatgc
     3061 aaagtctaaa aaggttaaga aaccaaaggc caagaaacca aaggaatcta ctgaatcatc
     3121 tcctgctcca ggtcaggaag ccgctgctac tgaaggtgcc acagaagata aacctaagaa
     3181 ggttaagaaa tcaaaggcta agaaggcaaa ggaatctgtt gaatcatctc ctgccgccac
     3241 tgaatctgtc tctgagaaga ctgcaaagaa atctaagaag ccaaaggcta agaaatctac
     3301 ttctcctgaa acaactgaag aaataactga agaatctacc gaatcaaagg aaaagaagac
     3361 aaagacaaag aaaccaaagg aaaagaagtc atctccatct actgccactt cgactgctgc
     3421 ttcaaaacca gttacttcca ttgcgggagt caccattcca ggcaagactt ttgatttgac
     3481 agatgttcct ccatctaaac ctgctccaaa gatggtcccg gaagcctata aattacaaac
     3541 tagagtggat tacggtacta agggtaccaa agtggacgtc ttgactaatc atatactact
     3601 atctgttggt gatgatgtcc ctcaagatga aagagcctct caattggatc catggtggaa
     3661 atctgcattt gtatatacct ataacattac ttttgccgtt ccacagagta attcaccacg
     3721 taaaggtcca gcgcctgctt tatccaaacc aaagaaatac gaattggtag aatctttgtt
     3781 tcacgaggat gaaaccttgt tcaaatataa agatcgtatc tccttcaatg gtgaagatac
     3841 tctatactct catgtcccat tggaggaatt cactttattt gatggttgtt gggatgtcag
     3901 taacaagcaa aagaaaagaa gacaagaagt tgttggtctc aacagtagag ccaaggaaat
     3961 taatgacttg gctgctcaag ttactttaaa atttgctgat aaagtcccat tgggtgatat
     4021 ttataaggct accacttcaa aggatccaga agaacaagaa aataagatgg ctaacgctga
     4081 taaggttgct ttattgtctt tgatgggtgt caaattctta aacacaaagg aacaaatctt
     4141 ccaacttaac ggtaataaat tctttatttt taatgaacac gcaattgcta ctccattcca
     4201 aattggtggg ttcttaatgc atgggttcac agtttcccta cgatatgctt acgggtccgt
     4261 tttgttaaat actgttaacg tctgcctccc atttgttaag tggactaaat atttaccagg
     4321 tgatgctaaa ttcaaggaaa atgaaaagac tcaatacagt ttgttagatt ggattattga
     4381 atgtatgcac caggccagtg ctcaaagagg tcaaaagcta agaggccctc catctgccaa
     4441 ggatatcaat tttttcattg acaagaatag agacattaaa gatctattga agggtttgaa
     4501 atgttacaga ccatacatta attattctgt taacccagat ggaactccaa aaccaccaaa
     4561 aaagatgcaa gctaagggta tcgttggttt cgtaagagag actccagatt ctatgaaatt
     4621 cagaacttta ccaagtaaca tggaaaagaa tggtgtacca aaaccaggtg aaaaggaaat
     4681 catggtcaca acaaccgcct actttgccaa gaagtatgac atcaaattga aatacccaga
     4741 tgttaaaatg gtgagtctag gtggttcgaa tgttgtccca gctgaatgtt taaccattgt
     4801 tccaggtcag aaattaaagg gtttggttta tgacgaaaaa gccgttattg atttcactgc
     4861 tttaagacca agtgaaaaat tcagagccat caccaacttg gcattacctg ccattaagag
     4921 agctttatca actgaagaag aaaatgctaa ggctccacat gattctggtt acactttcat
     4981 gaaggtccca tctcgtgtca ttgacgctcc tgtggttcaa ttcaagaaca ctacagtcac
     5041 atatgtggat aaaccatttg gtactaagaa tggtaagaat aatcatgaag aaactaaggg
     5101 taattggaat ttaaaggatc ataaattcat tactgttcca aaggaaccaa tgcacttgag
     5161 agctatcttt attaatgatt ctgataagtc tccaccagtt tctgtcatgg atgaattgaa
     5221 ggcctctttg agcaaattcg ctgaagatgt cgctgatgtc ggtgttaact ttgacgtatc
     5281 catggcccca attttaatta acaattttaa tgccccaata aagaaggtta ctggtggctt
     5341 tggtggtaga ggtggacgtg gtggtagagg cggccgtggt ggccgtggtg gtagaggagg
     5401 tcgtggtggt ttcggcggtg gccgtggtga aactacttat gaattaactc caggtgaaga
     5461 aaaactacgt cacttgttag ccaacgtccc agaaaagact tatgtcttat tcgtcttggg
     5521 tcgtggggat gactctgcta tttacaacag attgaaatat ttagctgatt tgacgtacgg
     5581 tgtcattaac aattgtgtta tttggaacaa gttcagaaag tgttccactc aatacaatgt
     5641 taatgtggta atgaagatga acttgaagtt agaaggtgct aaccactcct tatgtgcaga
     5701 agatataaac ttactgaagg atgaaaaatc aggattacca ttcatgattt taggtgctga
     5761 tgttacacat tatccagaaa aggatcagaa ttctatctct gcattggtgg gttcttttga
     5821 tgacaaattt gctcaattcc caggttcata tatgttacaa agtggtccag gtgaagaaat
     5881 aattgctggt atcggtaaca tggttctaca gagattgaaa ctgtatcaaa agcacaacaa
     5941 tggcaaacta cctccaaaga ttctattcta cagagatggt gtttctgaat cccaattctc
     6001 acaaattgtt caaattgaag ttaagggttt gaaacaagca ttgaagaaat tcggtagtga
     6061 attaaacaag ggtgtcaact ataacccatc tgtcaccaca atttgtgttg tcaagagaaa
     6121 tcaaattaga tttatgccac ttgagcaaaa tgctataaat gaaaagggtg aagttgctgc
     6181 cgttcaatca ttcgagaacg ttatgccagg tactgttgtt gatcgtggta tcacctctag
     6241 tgctcacttc gatttcttct tgcaatctca acaaccattg aagggtactg gtgttccatg
     6301 tcattactgg tgtatctacg acgaaaatca attcaattct gattacttgc aacaagttac
     6361 tcacgctttg tgttacttat tcggtagatc gagtacaagt attaaggtcg catctcctgt
     6421 ttattacgct gatttattgt gtgaacgtgg tgctgcattc ttcaaggcta actttgagct
     6481 tgctcaatac gaattttcta aggagaggaa gaacagagat gacgttatac cgaccggtaa
     6541 attactacaa cctgttcata agaatgtcac tgacatcatg tactacatat gactcgagtc
     6601 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa
     6661 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     6721 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     6781 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     6841 atttgcggcc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     6901 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     6961 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     7021 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     7081 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     7141 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     7201 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     7261 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     7321 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     7381 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     7441 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     7501 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggc
     7561 aagtgcacaa acaatactta aataaatact actcagtaat aacctatttc ttagcatttt
     7621 tgacgaaatt tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta
     7681 catcaacacc aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc
     7741 caccagctaa cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa
     7801 tcgagttcca atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa
     7861 acgaatgagg tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga
     7921 gtcttttaat aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat
     7981 gcagttggac gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt
     8041 gattacgaaa cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta
     8101 caagacttga aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac
     8161 atataatacc cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct
     8221 gcaagccgca aactttcacc aatggaccag aactacctgt gaaattaata acagacatac
     8281 tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc
     8341 ctccctcttg gccctctcct tttctttttt cgaccgaatt aattcttaat cggcaaaaaa
     8401 agaaaagctc cggatcaaga ttgtacgtaa ggtgacaagc tatttttcaa taaagaatat
     8461 cttccactac tgccatctgg cgtcataact gcaaagtaca catatattac gatgctgtct
     8521 attaaatgct tcctatatta tatatatagt aatgtcgttt atggtgcact ctcagtacaa
     8581 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     8641 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     8701 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
