LOCUS       sequence-106526-        4502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast ORF Puf6 in the Gateway entry vector pDONR221.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4502)
  AUTHORS   McCann KL, Charette JM, Vincent NG, Baserga SJ
  TITLE     A protein interaction map of the LSU processome.
  JOURNAL   Genes Dev. 2015 Apr 15;29(8):862-75. doi: 10.1101/gad.256370.114. 
            Epub 2015 Apr 15.
  PUBMED    25877921
REFERENCE   2  (bases 1 to 4502)
  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/gad.256370.114"; journalName: "Genes Dev"; date: 
            "2015-04-15- 15"; volume: "29"; issue: "8"; pages: "862-75"
COMMENT     Sequence Label: pDONR221 Puf6
FEATURES             Location/Qualifiers
     source          1..4502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2743..2762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2744..2762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2767..2783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2767..2783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2885..2904)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2896..3705
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2963..2982)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3852..4440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4341..4360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agatggcacc tttaaccaag aagacaaacg gaaaacgttc tgctaaagaa gttagtcatt
      721 cagaaaagaa attagccaaa aaacctagaa tttctattga ttcgtctgat gaagaatcag
      781 aactatccaa aaaggaagat gctgtatcct cttcatcaga tgatgatgat ttggatgatt
      841 taagtaccag tgactctgaa gcagaagaag aagctgacga attagatatt tccgatgaca
      901 gtgaggaaca tgaaaatgag aatgaagaga aagaaggaaa ggacaaatct gaaggaggtg
      961 aaaatggaaa tcacactgaa caaagaaaac tgctaaaaga aaggaaaatg caaagaaaat
     1021 ctggtaccca agtgcaacaa attaaatccg tctgggaaag attgcgtgtg aagaccccac
     1081 ctttaccaaa acagattcgt gaaaaacttt ccaacgaaat ctgggagcta tctaaggact
     1141 gtatcagtga tcttgtctta aagcatgatg cttcccgtat tgttcaaact ttagtcaagt
     1201 attcgtccaa agatcgtcgt gaacaaattg tcgatgcttt aaagggaaaa ttttatgttc
     1261 tagccacatc tgcgtacggt aagtatctat tggttaagtt actgcattat ggttcaagaa
     1321 gttcaaggca aactataata aatgaattac acggttcttt gagaaaatta atgagacatc
     1381 gtgagggtgc ctacgttgtt gaagatttat ttgttcttta tgccacacat gaacaaagac
     1441 aacaaatgat taaagagttt tggggctctg aatatgctgt atttagagaa acccacaaag
     1501 atttgaccat tgagaaagtt tgtgaaagta gcattgaaaa aaggaatatt atcgccagaa
     1561 acttgattgg taccattacc gcatcagtag aaaagggttc caccggcttt caaattctac
     1621 atgctgctat gagagaatat gtgaaaatag ccaacgaaaa ggaaatatct gaaatgatcg
     1681 aattgcttca tgaacaattc gctgaattgg tgcatactcc agaaggttca gacgttgctt
     1741 gtactttggt tgctagagct aacgccaagg aaagaaagtt aattttaaag gctttgaaaa
     1801 accacgccga aaaattgatc aaaaatgagt acggtaatat tgtcttcatt acaattctga
     1861 actgtgttga cgatacagtt ttagtattca agactttcag tcctactgtt aaggaacatt
     1921 tgcaagaatt cataattgat aagtttggta gaaggccttg gttatacata ctattaggtt
     1981 tggacggtaa gtatttctca cctatcgtca agaatgaatt attgagatac attgaactgt
     2041 ccaaagccac ctctaagaag gatcctttac aaagaaggca cgaacttttg tcaaaattcg
     2101 cccccatgtt tttaagcact atatcaaaag attattccag tatattgaca gagaatctag
     2161 gctgtcagtt cattgccgaa gttctaataa acgatgagtt gtacgcgcag ctgaatgaaa
     2221 aagaccaaga aaagtatcaa caggtcttaa acaacatttt aactactttt aagggtgaca
     2281 taaccgaaga agagcaccca atacatagag cattttctac tagactatta aaggcactaa
     2341 ttcaaggtgg taaatggaat aacaaagaaa aaaaggtcat accattaaaa aatgttcaag
     2401 gcttaggcgt tccatttgct gaaaagttat acgatgaaat tattgactcc tccaacttgt
     2461 tagaatggat taataacgct gatagttcat ttacaattgt tgcactgtat gaaacgttaa
     2521 aagatcaaaa agaagggaaa ccatttttga aggatttgag aggagttcag agtaaaatca
     2581 ccactgatga aagtaacaaa ggctctcagc ttttggctaa attgttaaaa agctttcttg
     2641 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     2701 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2761 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2821 attgcacaag ataaaataat atcatcatga acaataaaac tgtctgctta cataaacagt
     2881 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2941 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     3001 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3061 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     3121 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     3181 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     3241 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     3301 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     3361 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     3421 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     3481 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3541 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3601 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3661 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3721 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3781 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3841 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3901 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3961 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4021 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4081 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4141 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4201 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4261 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4321 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4381 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4441 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4501 tt
//
