LOCUS       sequence-45191-e        5919 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 5919)
  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 5919)
  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: pRS403-PGAL1-hpSC_URA3
FEATURES             Location/Qualifiers
     source          1..5919
                     /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"
     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        2316..2757
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2316..2433
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2728..2751
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      3560..3807
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3576..3594)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3852..3871)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3853..3871)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3881..3898)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3881..3897)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3890..3912)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4042..4497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4129..4148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4339..4360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4757..5416)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5417..5603)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5750..5769)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5869..5891
                     /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 tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc
     2281 accgcggtgg cggccgcggg gatgatccac tagtacggat tagaagccgc cgagcgggtg
     2341 acagccctcc gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg
     2401 aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct
     2461 tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac
     2521 gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg
     2581 taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg
     2641 cataaccact ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg
     2701 taataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa
     2761 aaaccccgga tcggactact agcagctgta aaggcattat ccgccaagta caatttttta
     2821 ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     2881 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     2941 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     3001 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     3061 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     3121 atgggtgctc gagaaaatca ggtgtgttgg aacttttttc aaaccttact aaacattgaa
     3181 actaattggt aaagatagag gaattccacc catgtctctt tgagcaataa agccgataac
     3241 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     3301 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     3361 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     3421 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     3481 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     3541 ctttatctag agggccgcat catgtaatta gttatgtcac gcttacattc acgccctccc
     3601 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     3661 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     3721 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     3781 ttgggacgct cgaaggcttt aatttgcaag ctgcggcgtc gacctcgagg gggggcccgg
     3841 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     3901 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3961 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4021 gcctgaatgg cgaatggcgc gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4081 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4141 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4201 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4261 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4321 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4381 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4441 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcct
     4501 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag atccgtcgag
     4561 ttcaagagaa aaaaaaagaa aaagcaaaaa gaaaaaagga aagcgcgcct cgttcagaat
     4621 gacacgtata gaatgatgca ttaccttgtc atcttcagta tcatactgtt cgtatacata
     4681 cttactgaca ttcataggta tacatatata cacatgtata tatatcgtat gctgcagctt
     4741 taaataatcg gtgtcactac ataagaacac ctttggtgga gggaacatcg ttggtaccat
     4801 tgggcgaggt ggcttctctt atggcaaccg caagagcctt gaacgcactc tcactacggt
     4861 gatgatcatt cttgcctcgc agacaatcaa cgtggagggt aattctgcta gcctctgcaa
     4921 agctttcaag aaaatgcggg atcatctcgc aagagagatc tcctactttc tccctttgca
     4981 aaccaagttc gacaactgcg tacggcctgt tcgaaagatc taccaccgct ctggaaagtg
     5041 cctcatccaa aggcgcaaat cctgatccaa acctttttac tccacgcgcc agtagggcct
     5101 ctttaaaagc ttgaccgaga gcaatcccgc agtcttcagt ggtgtgatgg tcgtctatgt
     5161 gtaagtcacc aatgcactca acgattagcg accagccgga atgcttggcc agagcatgta
     5221 tcatatggtc cagaaaccct atacctgtgt ggacgttaat cacttgcgat tgtgtggcct
     5281 gttctgctac tgcttctgcc tctttttctg ggaagatcga gtgctctatc gctaggggac
     5341 caccctttaa agagatcgca atctgaatct tggtttcatt tgtaatacgc tttactaggg
     5401 ctttctgctc tgtcatcttt gccttcgttt atcttgcctg ctcatttttt agtatattct
     5461 tcgaagaaat cacattactt tatataatgt ataattcatt atgtgataat gccaatcgct
     5521 aagaaaaaaa aagagtcatc cgctagggga aaaaaaaaaa tgaaaatcat taccgaggca
     5581 taaaaaaata tagagtgtac tagaggaggc caagagtaat agaaaaagaa aattgcggga
     5641 aaggactgtg ttatgacttc cctgactaat gccgtgttca aacgatacct ggcagtgact
     5701 cctagcgctc accaagctct taaaacggga atttatggtg cactctcagt acaatctgct
     5761 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5821 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5881 tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
