LOCUS       sequence-138501-        7474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Uracil selection; Expression of PP7-2x-yeGFP fusion protein under 
            the ADE3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7474)
  AUTHORS   Saroufim MA, Bensidoun P, Raymond P, Rahman S, Krause MR, Oeffinger 
            M, Zenklusen D
  TITLE     The nuclear basket mediates perinuclear mRNA scanning in budding 
            yeast.
  JOURNAL   J Cell Biol. 2015 Dec 21;211(6):1131-40. doi: 10.1083/jcb.201503070.
  PUBMED    26694838
REFERENCE   2  (bases 1 to 7474)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2015-12-21- 21"; volume: "211"; 
            issue: "6"; pages: "1131-40"
COMMENT     Sequence Label: pDZ536 pURA ADE3p PP7-PS-2x-yeGFP
FEATURES             Location/Qualifiers
     source          1..7474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1022..1732
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1742..2455
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      2465..2712
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2481..2499)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(2734..2752)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2773..2789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2773..2789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2786..2808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2797..2813
                     /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)."
     promoter        complement(join(2821..2827,2828..2845,2846..2851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2866..2887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3004..3021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3175..3763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3255..3274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3934..4725,4726..4794))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4557..4576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4795..4899)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4936..5439
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5481..5499
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5537..5559)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5659..5678
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5698..5918
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5919..6722
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(6851..7306)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6988..7009)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7200..7219
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7432..7454
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7446..7463
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7447..7463
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7470..7474,1..15)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(7470..7474,1..14)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgactcacta tagggcgaat tggagctctc caggtagtgg atgtgcgagg ctcagattat
       61 atggggggcc atatcaagga cggatggcac tacgcctatt cgcgccttaa gcaggatccg
      121 gagtacttac gtgagctaaa gcacagattg ttggaaaagc aagcggacgg ccgcggagcg
      181 ctgaacgtga ttttccattg tatgttatcg cagcagcgcg gaccgtctgc agcgatgctg
      241 ctgcttcggt cgcttgacac ggcggaactt tctcgctgtc gtctatgggt gttgcgtggg
      301 gggttctcgc gctggcaatc cgtatacggt gacgacgaga gcgttacggc gggttaccta
      361 cccgatctgt ggcgttgaca ggtttacaca atcgcacgtg atcatatatt tgccatgact
      421 cctcccagtg acaattttgt tcttttttcc tctttatcgc tttcgtacta tggtcagtca
      481 ttcattcatt atatacgcgc tctccataac ccgtaacttt ttattatata tagactcgtt
      541 tacaatacaa cgatagcgat accattcaat tgaagttgtg agaccaggta acgagacgaa
      601 cacaacttta caagtctcta gaatgtccaa aaccatcgtt ctttcggtcg gcgaggctac
      661 tcgcactctg actgagatcc agtccaccgc agaccgtcag atcttcgaag agaaggtcgg
      721 gcctctggtg ggtcggctgc gcctcacggc ttcgctccgt caaaacggag ccaagaccgc
      781 gtatcgcgtc aacctaaaac tggatcaggc ggacgtcgtt gattccggac ttccgaaagt
      841 gcgctacact caggtatggt cgcacgacgt gacaatcgtt gcgaatagca ccgaggcctc
      901 gcgcaaatcg ttgtacgatt tgaccaagtc cctcgtcgcg acctcgcagg tcgaagatct
      961 tgtcgtcaac cttgtgccgc tgggccgtgg atcccccggg ctgcaggaat tcgatatcgt
     1021 gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
     1081 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
     1141 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
     1201 cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata tgaaacaaca
     1261 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     1321 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     1381 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     1441 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
     1501 caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca
     1561 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     1621 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     1681 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aagatatcgt
     1741 gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
     1801 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
     1861 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
     1921 cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata tgaaacaaca
     1981 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     2041 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     2101 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     2161 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
     2221 caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca
     2281 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     2341 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     2401 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aataactggt
     2461 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     2521 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     2581 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     2641 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     2701 gctttaattt gcggccggta ccagcttttg ttccctttag tgagggttaa tttcgagctt
     2761 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2821 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2881 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2941 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3601 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactgcccg tcgtgtagat
     4021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4321 agttacatga tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt
     4381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4921 acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
     4981 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
     5041 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
     5101 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
     5161 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
     5221 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
     5281 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
     5341 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
     5401 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa accattatta
     5461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     5521 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5581 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5641 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cacgcttttc
     5701 aattcaattc atcatttttt ttttattctt ttttttgatt tcggtttcct tgaaattttt
     5761 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     5821 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     5881 tgcacagaac aaaaacctgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     5941 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     6001 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     6061 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     6121 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     6181 tcttcgaaga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     6241 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     6301 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     6361 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     6421 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     6481 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     6541 acaagggaga cgcattgggt caacagtata gaaccgtgga tgatgtggtc tctacaggat
     6601 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     6661 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     6721 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     6781 aattatatca gttattaccc tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6841 ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6901 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6961 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     7021 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7081 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7141 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7201 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7261 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     7321 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     7381 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     7441 gacgttgtaa aacgacggcc agtgaattgt aata
//
