LOCUS       sequence-122741-        7256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TCP4 expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7256)
  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 7256)
  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: pIR195
FEATURES             Location/Qualifiers
     source          1..7256
                     /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(1853..1877)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1900..1941
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1951..1968
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2002..2249
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2018..2036)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2326..2343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2497..3085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2577..2596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3256..4047,4048..4116))
                     /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(4212..5015)
                     /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(5016..5236)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5835..6715
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6784..7239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7133..7152
                     /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 ttcgcccttc ccggatccat gtctgacgac caattccatc acccgccgcc tccttcttca
      601 atgaggcacc gttctacgtc ggatgcggcg gacggcggct gcggcgagat tgttgaggtg
      661 caaggtggtc acattgttcg gtctaccgga agaaaagacc gccacagcaa agtctgcacg
      721 gctaaagggc cacgtgaccg gcgcgtgaga ctctctgctc acacggcgat tcagttttac
      781 gatgttcaag acaggcttgg tttcgaccga cctagcaaag ccgttgattg gcttatcaaa
      841 aaggctaaga cttccattga cgagctcgct gagcttcctc cctggaatcc cgccgatgca
      901 attcgcctag ccgctgctaa cgctaaaccc agaagaacca ccgccaaaac ccaaatctct
      961 ccgtctccgc caccgccgca acagcaacaa caacaacaac agcttcagtt cggtgttggc
     1021 ttcaacggag gaggagcaga gcatccgagt aacaacgagt cgagttttct cccgccgtca
     1081 atggattcag attcgatagc tgacactata aagtcgtttt ttccggtgat tggctcttca
     1141 acggaggctc cttcgaatca taaccttatg cacaactatc atcatcagca tccgccggat
     1201 ttgctttctc gaactaatag ccaaaaccaa gatctccgtc tctcgctgca atcgttcccg
     1261 gatggtccac cgtcgcttct gcaccaccaa catcaccacc acacctctgc ttccgcctcc
     1321 gagcctactc tgttctacgg acagagcaat ccgttagggt ttgacacatc gagttgggag
     1381 cagcagtcgt cggaattcgg aaggattcag agactagtgg cttggaacag cggcggtggc
     1441 ggcggagcaa ccgatacagg aaacggagga gggtttctgt tcgctcctcc tactccttca
     1501 acgacgtcgt ttcagccagt tcttggccaa agccaacagc tttattctca gaggggtccc
     1561 ttgcaaagta gctacagtcc catgatccgt gcttggtttg atcctcacca tcatcaccaa
     1621 tccatctcca ccgacgatct caaccaccac catcaccttc ctccaccggt tcaccaatca
     1681 gcaatccccg gaatcggatt cgcctcaggt gaattctctt cgggttttcg cataccagca
     1741 cggtttcagg gccaagaaga ggagcagcac gacggtctca ctcacaagcc gtcctctgct
     1801 tcctctattt ctcgccattg aaaaggccct gtcgacggga agggcgaatt cgacccagct
     1861 ttcttgtaca aagtggttcg atctagaggg cccttcgaag gtaagcctat ccctaaccct
     1921 ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc
     1981 cgctgatcct agagggccgc atcatgtaat tagttatgtc acgcttacat tcacgccctc
     2041 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     2101 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     2161 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     2221 ttttgggacg ctcgaaggct ttaatttgca agctgcggcc ctgcattaat gaatcggcca
     2281 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2461 gcccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggagcg
     3361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3541 taatagtttg cgcaacgttg ttggcattgc tacaggcatc gtggtgtcac tctcgtcgtt
     3601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aatagtgtat cacatagcag
     3901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatgggtaat
     4141 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     4201 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     4261 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     4321 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     4381 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     4441 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     4501 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     4561 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     4621 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     4681 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     4741 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     4801 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     4861 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     4921 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     4981 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     5041 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     5101 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     5161 attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     5221 gatgaattga attgaaaagc tagcttatcg atgataagct gtcaaagatg agaattaatt
     5281 ccacggacta tagactatac tagatactcc gtctactgta cgatacactt ccgctcaggt
     5341 ccttgtcctt taacgaggcc ttaccactct tttgttactc tattgatcca gctcagcaaa
     5401 ggcagtgtga tctaagattc tatcttcgcg atgtagtaaa actagctaga ccgagaaaga
     5461 gactagaaat gcaaaaggca cttctacaat ggctgccatc attattatcc gatgtgacgc
     5521 tgcagcttct caatgatatt cgaatacgct ttgaggagat acagcctaat atccgacaaa
     5581 ctgttttaca gatttacgat cgtacttgtt acccatcatt gaattttgaa catccgaacc
     5641 tgggagtttt ccctgaaaca gatagtatat ttgaacctgt ataataatat atagtctagc
     5701 gctttacgga agacaatgta tgtatttcgg ttcctggaga aactattgca tctattgcat
     5761 aggtaatctt gcacgtcgca tccccggttc attttctgcg tttccatctt gcacttcaat
     5821 agcatatctt tgttaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     5881 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     5941 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     6001 cgcgacgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     6061 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     6121 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     6181 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     6241 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     6301 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     6361 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     6421 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     6481 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     6541 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     6601 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     6661 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatgggaa
     6721 gctccacccc ggttgataat cagaaaagcc ccaaaaacag gaagattgta taagcaaata
     6781 tttaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6841 cattttttaa cgaatagccc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6901 agatagggtt gagtgttgtt ccagtttcca acaagagtcc actattaaag aacgtggact
     6961 ccaacgtcaa agggcgaaaa agggtctatc agggcgatgg cccactacgt gaaccatcac
     7021 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcagt aaatcggaag ggtaaacgga
     7081 tgcccccatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     7141 aagcgaaagg agcgggggct agggcggtgg gaagtgtagg ggtcacgctg ggcgtaacca
     7201 ccacacccgc cgcgcttaat ggggcgctac agggcgcgtg gggatgatcc actagt
//
