LOCUS       sequence-162947-        5279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid can be used as a donor for subcloning into Gateway 
            destination vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5279)
  AUTHORS   Chymkowitch P, Nguea AP, Aanes H, Koehler CJ, Thiede B, Lorenz S, 
            Meza-Zepeda LA, Klungland A, Enserink JM
  TITLE     Sumoylation of Rap1 mediates the recruitment of TFIID to promote 
            transcription of ribosomal protein genes.
  JOURNAL   Genome Res. 2015 Jun;25(6):897-906. doi: 10.1101/gr.185793.114. Epub
            2015 Mar 23.
  PUBMED    25800674
REFERENCE   2  (bases 1 to 5279)
  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: 
            "10.1101/gr.185793.114"; journalName: "Genome Res"; date: "2015-06";
            volume: "25"; issue: "6"; pages: "897-906"
COMMENT     Sequence Label: pDONR221-rap1-K651R
FEATURES             Location/Qualifiers
     source          1..5279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2679..2778)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2795..2814)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2796..2814)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2819..2835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2819..2835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3266..4057
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4150..4738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4639..4658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4892..4909
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5068..5095)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5187..5273)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaga
      181 aagctggcta caaaatgtct agtccagatg attttgaaac tgcaccagca gaatatgttg
      241 atgcattgga tccgagtatg gtcgttgttg acagtggttc tgctgcagtc acagcgccat
      301 cggattcagc cgctgaggta aaagctaatc aaaacgaaga aaacactggt gctactgctg
      361 ctgaaactag tgagaaagtg gatcagacgg aggtcgaaaa gaaagatgat gatgatacca
      421 ctgaagtcgg tgtaacaact accacgcctt ccattgctga tactgctgct accgctaaca
      481 ttgcttcgac atcaggtgca tctgttactg aacctactac tgatgacacc gctgctgatg
      541 agaaaaagga acaagtaagt ggtcctcctc tgtcaaatat gaaattctat cttaatcgcg
      601 acgcggatgc gcatgactct ttaaatgata ttgatcaatt agcgcgtctg attagagcaa
      661 acggtggtga agtgctagac tccaagccaa gagaatcaaa agagaatgtt ttcatcgtgt
      721 caccctataa tcataccaat ttaccaacgg taactccgac ttatattaag gcatgttgtc
      781 aaagcaactc gctactgaac atggaaaatt atttagtacc atacgataac tttagagaag
      841 ttgtcgatag taggttgcag gaggaatcgc attctaatgg tgtagataat agcaatagca
      901 atagcgataa caaggattct atcaggccca aaacagaaat aatctctacg aatactaatg
      961 gcgctacgga agactcaacc agcgaaaaag ttatggtaga cgctgaacaa caagctagat
     1021 tacaagagca agctcagctc cttcgtcaac atgtaagcag caccgcatca atcacaagcg
     1081 gagggcacaa tgacttggtc cagatcgaac aacctcaaaa agatacctcc aataataata
     1141 atagcaacgt caacgacgaa gacaatgatc ttttgacaca ggacaacaac cctcaaacag
     1201 cagatgaggg gaatgcatct tttcaagcac aaaggtccat gatttcgaga ggcgctttgc
     1261 cctcccacaa taaagcttct tttacagatg aggaagatga gtttattttg gatgttgtga
     1321 gaaaaaatcc aaccaggcgt acaacacata ctctttacga tgaaatatcc cattatgtgc
     1381 ctaaccacac gggtaattct attaggcacc gatttagagt ctatctttcc aaaagactag
     1441 agtacgttta tgaggttgac aagtttggta aattggttag agacgacgat ggaaatctga
     1501 taaagactaa agttttgcca ccatcaatta aaaggaaatt ttcagcagat gaagattaca
     1561 ctttggcaat cgcggtcaag aagcagtttt atcgtgattt gtttcaaatc gaccctgata
     1621 ctggaagatc acttatcaca gatgaggata cacccactgc tatagcgagg aggaatatga
     1681 caatggatcc aaatcatgtg ccagggagtg aacccaactt tgcagcttac agaacccaaa
     1741 gccgtagggg accgattgca cgagaatttt tcaagcattt tgccgaagag catgcagcac
     1801 atactgagaa tgcttggaga gataggttta ggaagtttct tctcgcttac ggcattgatg
     1861 attatattag ttattatgaa gctgaaaaag ctcagaatag agagccggaa ccgatgaaga
     1921 acctcaccaa tagacccaag aggcctggcg ttcccactcc tggcaattat aactctgccg
     1981 ccaagagggc aagaaattat agttctcaaa gaaatgttca gcctactgcc aatgcagcgt
     2041 ccgctaatgc tgctgccgct gctgctgccg ctgcttccaa ctcttacgct ataccagaaa
     2101 acgaattact ggacgaagat accatgaatt tcatttccag tttaagaaat gatctatcca
     2161 atatttcaaa tagtttgccc tttgagtatc cacacgagat tgcggaagct ataagaagcg
     2221 atttttcgaa tgaagatata tatgataata ttgatcctga taccataagc tttccaccaa
     2281 aaattgcaac aacagatctt ttcctgccat tgttctttca ttttggcagt acgagacaat
     2341 ttatggataa acttcatgaa gttatttcag gagattatga gccatcacag gctgaaaaac
     2401 tggtacagga tctttgcgat gaaactggta tacgtaaaaa tttcagtacg agcatattga
     2461 cgtgtttatc cggggatctg atggtctttc ctcgctattt cttgaacatg tttaaggaca
     2521 atgttaatcc tcctcccaac gttcctggta tttggacaca tgatgacgac gaatcgctaa
     2581 aaagcaatga tcaagaacaa ataaggaaac tggttaaaaa gcatggaact ggtagaatgg
     2641 aaatgaggaa aagatttttt gagaaggacc tgttatgaac ccagctttct tgtacaaagt
     2701 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2761 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2821 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2881 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
     2941 gctgctgccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     3001 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     3061 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     3121 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     3181 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     3241 tcgccctaaa acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca
     3301 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     3361 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     3421 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     3481 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     3541 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     3601 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     3661 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     3721 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     3781 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     3841 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     3901 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     3961 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     4021 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     4081 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4261 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     4561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4861 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4921 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     4981 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
     5041 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     5101 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     5161 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     5221 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcct
//
