LOCUS       sequence-112047-        8422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CUC2 expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8422)
  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 8422)
  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: JW624
FEATURES             Location/Qualifiers
     source          1..8422
                     /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      2456..2503
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2530..2717
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2585..2609)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2741..2757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2741..2757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2754..2776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2765..2781
                     /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(2789..2795,2796..2813,2814..2819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2972..2989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3143..3731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3223..3242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4060..4107
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4132..4151
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4340..5134)
                     /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(4451..4470)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5061..5080
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5135..5239)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5266..6608
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6229..6276)
                     /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     6649..6667
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6705..6727)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6827..6846
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6867..7148
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7149..7823
                     /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(7926..8381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8063..8084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8275..8294
                     /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 attcatggac attccgtatt
     1321 accactacga ccatggcgga gacagccaat atcttccacc gggtttcagg tttcatccca
     1381 cggacgaaga gctcatcact cattaccttc tccgcaaagt cctcgacggt tgcttctcaa
     1441 gccgtgccat cgcagaagtt gatctcaaca agtgtgagcc ttggcaactt cccgggagag
     1501 ctaagatggg agagaaagaa tggtacttct ttagcctccg tgaccggaag tatccgacgg
     1561 gactgagaac taacagagca actgaggctg gttactggaa agctaccgga aaagacagag
     1621 agatctttag ttcaaagact tgtgcacttg ttgggatgaa gaagactctt gtcttttaca
     1681 aaggaagagc tccgaaagga gagaagagta attgggttat gcatgaatat cgtcttgaag
     1741 gcaaattctc ttaccatttc atctcaagaa gctccaagga tgaatgggtg atctctaggg
     1801 ttttccagaa aaccacttta gctagcaccg gagccgtctc cgaaggagga ggaggaggag
     1861 gagcaactgt gagcgtaagc agcggtactg gtccatctaa aaagacgaaa gtaccctcaa
     1921 caatctcaag aaactatcaa gaacaaccaa gctctccttc ctccgtctca ctcccacctc
     1981 tcctggatcc gaccactacc ctcggctaca ccgacagcag ttgctcctac gacagccgta
     2041 gcaccaacac aaccgtcaca gccagcgcaa taaccgagca cgtgtcctgt ttctccactg
     2101 tccctactac tactacggcc ttgggcttag acgttaactc attcagccgt cttccaccgc
     2161 cgctagggtt tgactttgac ccttttcctc gtttcgtttc tagaaacgtc tcgactcaat
     2221 ctaacttcag atcgttccaa gaaaacttca atcaatttcc ttactttgga tcgtcttctg
     2281 catcgactat gacctccgcc gttaatctgc cttctttcca aggcggcgga ggcgtctccg
     2341 ggatgaatta ctggctaccg gcgactgccg aagagaatga gtcaaaggtc ggtgtgcttc
     2401 atgctggact tgactgtatt tggaactact gagtcgacct gcagcggccg cataactagc
     2461 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgcgcgctt gcagccaagc
     2521 taattccggg cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     2581 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     2641 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt
     2701 gaccacacct ctaccggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2761 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2821 cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt
     2881 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2941 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3001 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3061 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3121 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3181 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3241 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3301 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3361 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3421 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3481 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3541 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3601 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3661 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3721 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3781 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3841 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     3901 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     3961 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     4021 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     4081 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     4141 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     4201 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     4261 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggttcatt
     4321 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     4381 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     4441 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     4501 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     4561 aggcatcgtc atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg
     4621 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     4681 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     4741 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     4801 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     4861 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     4921 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     4981 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     5041 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     5101 gagaacctgc gtgcaatcca tcttgttcaa tcatactctt cctttttcaa tattattgaa
     5161 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5221 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaacga agcatctgtg
     5281 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     5341 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     5401 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     5461 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     5521 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     5581 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     5641 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     5701 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     5761 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     5821 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     5881 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     5941 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     6001 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     6061 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     6121 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     6181 tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga agttcctata
     6241 ctttctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct
     6301 tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct
     6361 gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa
     6421 atgcgtactt atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat
     6481 attatcccat tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct
     6541 atatgctgcc actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat
     6601 attggatcat attaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6661 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6721 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6781 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6841 attgtactga gagtgcacca tagatcaacg acattactat atatataata taggaagcat
     6901 ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag
     6961 tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc
     7021 ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag
     7081 agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag
     7141 cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg
     7201 gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta
     7261 ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa
     7321 gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc
     7381 gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc
     7441 aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta
     7501 aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc
     7561 attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga
     7621 actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag
     7681 ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg
     7741 ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt
     7801 tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt
     7861 gcacttgccg atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7921 tcaggaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     7981 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     8041 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     8101 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     8161 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     8221 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     8281 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     8341 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     8401 cgcaactgtt gggaagggcg at
//
