LOCUS       sequence-122730-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SPL9 expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  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 7094)
  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: pIR196
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     414..437
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     475..494
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    511..535
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1691..1715)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1738..1779
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1789..1806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1840..2087
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1856..1874)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2164..2181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2415..2434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3094..3885,3886..3954))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(4050..4853)
                     /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"
     promoter        complement(4854..5074)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5673..6553
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6622..7077)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6971..6990
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagctatcaa acaagtttgt acaaaaaagc aggctccgaa
      541 ttcgccctta tggagatggg ttccaactcg ggtccgggtc atggtccggg tcaggcagag
      601 tcgggtggtt cctccactga gtcatcctct ttcagtggag ggctcatgtt tggccagaag
      661 atctacttcg aggacggtgg tggtggatcc gggtcttctt cctcaggtgg tcgttcaaac
      721 agacgtgtcc gtggaggcgg gtcgggtcag tcgggtcaga taccaaggtg ccaagtggaa
      781 ggttgtggga tggatctaac caatgcaaaa ggttattact cgagacaccg agtttgtgga
      841 gtgcactcta aaacacctaa agtcactgtg gctggtatcg aacagaggtt ttgtcaacag
      901 tgcagcaggt ttcatcagct tccggaattt gacctagaga aaaggagttg ccgcaggaga
      961 ctcgctggtc ataatgagcg acgaaggaag ccacagcctg cgtctctctc tgtgttagct
     1021 tctcgttacg ggaggatcgc accttcgctt tacgaaaatg gtgatgctgg aatgaatgga
     1081 agctttcttg ggaaccaaga gataggatgg ccaagttcaa gaacattgga tacaagagtg
     1141 atgaggcggc cagtgtcgtc accgtcatgg cagatcaatc caatgaatgt atttagtcaa
     1201 ggttcagttg gtggaggagg gacaagcttc tcatctccag agattatgga cactaaacta
     1261 gagagctaca agggaattgg cgactcaaac tgcgccttaa gcttgttaag caatccacat
     1321 caaccacatg acaacaacaa caacaacaac aacaacaaca acaacaacaa caatacatgg
     1381 cgagcttctt caggttttgg cccgatgacg gttacaatgg ctcaaccacc acctgcacct
     1441 agccagcatc agtatctgaa cccgccttgg gtattcaagg acaatgataa tgatatgtct
     1501 cctgttttga atttaggtcg atacaccgag ccagataatt gtcagataag tagtggcacg
     1561 gcaatgggtg agttcgagtt atctgatcac catcatcaaa gtaggagaca gtacatggaa
     1621 gatgagaaca caagggctta tgactcttct tctcaccata ccaactggtc tctctgaaag
     1681 ggcgaattcg acccagcttt cttgtacaaa gtggttcgat ctagagggcc cttcgaaggt
     1741 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat
     1801 caccattgag tttaaacccg ctgatcctag agggccgcat catgtaatta gttatgtcac
     1861 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     1921 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     1981 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     2041 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctgcggccct
     2101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2281 agcaaaaggc cagcaaaagc ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2761 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3061 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3181 aactacgata cgggagcgct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3301 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt ggcattgcta caggcatcgt
     3421 ggtgtcactc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3541 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3601 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3721 tagtgtatca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3781 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3961 cctttttcaa tgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     4021 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     4081 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     4141 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     4201 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     4261 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     4321 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     4381 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     4441 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     4501 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     4561 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     4621 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     4681 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     4741 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     4801 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     4861 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     4921 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     4981 tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa
     5041 aaagaataaa aaaaaaatga tgaattgaat tgaaaagcta gcttatcgat gataagctgt
     5101 caaagatgag aattaattcc acggactata gactatacta gatactccgt ctactgtacg
     5161 atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt tgttactcta
     5221 ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac
     5281 tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat
     5341 tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac
     5401 agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga
     5461 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     5521 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     5581 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     5641 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     5701 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5761 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     5821 tgtaaaacaa aaatgcaacg cgacgagagc gctaattttt caaacaaaga atctgagctg
     5881 catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac
     5941 ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta
     6001 gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact
     6061 gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa
     6121 agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca
     6181 agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca
     6241 gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt
     6301 ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc
     6361 tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa
     6421 aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat
     6481 atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag
     6541 cggtattcgc aatgggaagc tccaccccgg ttgataatca gaaaagcccc aaaaacagga
     6601 agattgtata agcaaatatt taaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     6661 atttttgtta aatcagctca ttttttaacg aatagcccga aatcggcaaa atcccttata
     6721 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttccaac aagagtccac
     6781 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaag ggtctatcag ggcgatggcc
     6841 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcagtaa
     6901 atcggaaggg taaacggatg cccccattta gagcttgacg gggaaagccg gcgaacgtgg
     6961 cgagaaagga agggaagaaa gcgaaaggag cgggggctag ggcggtggga agtgtagggg
     7021 tcacgctggg cgtaaccacc acacccgccg cgcttaatgg ggcgctacag ggcgcgtggg
     7081 gatgatccac tagt
//
