LOCUS       sequence-122702-        8327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CUC3 expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8327)
  AUTHORS   Rubio-Somoza I, Zhou CM, Confraria A, Martinho C, von Born P, 
            Baena-Gonzalez E, Wang JW, Weigel D
  TITLE     Temporal control of leaf complexity by miRNA-regulated licensing of 
            protein complexes.
  JOURNAL   Curr Biol. 2014 Nov 17;24(22):2714-9. doi: 
            10.1016/j.cub.2014.09.058. Epub 2014 Nov 6.
  PUBMED    25448000
REFERENCE   2  (bases 1 to 8327)
  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: 
            "10.1016/j.cub.2014.09.058"; journalName: "Curr Biol"; date: 
            "2014-11-17- 17"; volume: "24"; issue: "22"; pages: "2714-9"
COMMENT     Sequence Label: JW617
FEATURES             Location/Qualifiers
     source          1..8327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     1213..1232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      2361..2408
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2435..2622
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2490..2514)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2646..2662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2646..2662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2659..2681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2670..2686
                     /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(2694..2700,2701..2718,2719..2724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2877..2894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3048..3636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3128..3147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3965..4012
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4037..4056
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4245..5039)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4356..4375)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4966..4985
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5040..5144)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5171..6513
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6134..6181)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6554..6572
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6610..6632)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6732..6751
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6772..7053
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7054..7728
                     /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"
     rep_origin      complement(7831..8286)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7968..7989)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8180..8199
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaaa tgatggaaga
      421 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg
      481 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct
      541 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcgccctt atgatgcttg
     1321 cggtggaaga tgtgttaagc gaactcgccg gagaagaaag gaacgagaga ggattgccac
     1381 ctggcttccg gtttcacccg acggacgaag agctcattac cttctactta gcttccaaaa
     1441 tcttccatgg tggtctctcc ggcattcaca tttccgaagt tgatctcaac cgctgtgaac
     1501 cttgggagct accagaaatg gcgaagatgg gagagagaga gtggtacttt tatagtctaa
     1561 gggacaggaa atatccgaca ggtttgagga ctaacagagc aactactgct ggatactgga
     1621 aagctaccgg caaagataag gaagtcttct ccggcggagg aggacagctt gttgggatga
     1681 agaagacgtt ggtgttctac aaaggtaggg ctccacgtgg cctcaagact aagtgggtca
     1741 tgcatgagta tcgcctcgaa aacgaccatt cacaccgcca cacgtgtaag gaggaatggg
     1801 tgatttgcag agtgttcaat aaaacaggag acagaaaaaa tgttggatta atccataacc
     1861 aaatcagcta ccttcataac cattcactct caacaacaca tcatcatcat catgaagcct
     1921 tacctttgct tatagaacct tccaacaaaa ccctaaccaa cttcccatca ctactctacg
     1981 atgatccaca ccaaaactac aataataaca acttccttca tggatcatca ggccacaaca
     2041 tcgacgagct caaagcctta atcaaccctg tcgtctctca gctcaacggt atcatctttc
     2101 cttcagggaa caacaacaac gacgaagacg acttcgactt taacctcggc gtgaaaacag
     2161 agcagtcttc gaacggtaac gaaattgacg tacgagatta cttggagaac cctctgtttc
     2221 aggaagcgag ttatggtctg ttgggttttt cgtcttctcc tggacctctt cacatgctac
     2281 tagattctcc atgtccttta ggattccagc tgtagaaggg cgaattcccg gggatccgtc
     2341 gacctgcagc ggccgcataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2401 gttttttgcg cgcttgcagc caagctaatt ccgggcgaat ttcttatgat ttatgatttt
     2461 tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt
     2521 tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca
     2581 ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgcaag cttggcgtaa
     2641 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2701 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta
     2761 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2821 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2881 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2941 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3781 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     3841 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     3901 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     3961 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     4021 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     4081 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     4141 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     4201 ttgggcgtcg cttggtcggt tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     4261 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     4321 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     4381 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     4441 ttttccacca tgatattcgg caagcaggca tcgtcatggg tcacgacgag atcctcgccg
     4501 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     4561 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     4621 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     4681 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     4741 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     4801 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     4861 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4921 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4981 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcata
     5041 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5101 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5161 gtgccacctg aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     5221 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     5281 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     5341 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     5401 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     5461 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     5521 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     5581 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     5641 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     5701 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     5761 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     5821 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     5881 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     5941 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     6001 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     6061 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt
     6121 tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt
     6181 caaagcgttt ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct
     6241 cactgttcac gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa
     6301 cggcatagtg cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga
     6361 aaggtagtct agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc
     6421 ttcagcacta ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc
     6481 cttcaatgct atcatttcct ttgatattgg atcatattaa gaaaccatta ttatcatgac
     6541 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     6601 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     6661 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     6721 gcttaactat gcggcatcag agcagattgt actgagagtg caccatagat caacgacatt
     6781 actatatata taatatagga agcatttaat agaacagcat cgtaatatat gtgtactttg
     6841 cagttatgac gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac
     6901 cttacgtaca atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc
     6961 gaaaaaagaa aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta
     7021 tacgtgatta agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt
     7081 tctggtccat tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta
     7141 gattccgatg ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac
     7201 ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact
     7261 ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc
     7321 aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa
     7381 gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta
     7441 ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt
     7501 gggacaggtg aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc
     7561 cccgaaagct tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg
     7621 cttagattaa atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta
     7681 aaagactcta acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag
     7741 tagtatttat ttaagtattg tttgtgcact tgccgatcta tgcggtgtga aataccgcac
     7801 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     7861 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     7921 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     7981 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     8041 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     8101 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     8161 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     8221 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     8281 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgat
//
