LOCUS       sequence-115919-        5073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast ORF Spb1 in the Gateway entry vector pDONR221.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  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 5073)
  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 Spb1
FEATURES             Location/Qualifiers
     source          1..5073
                     /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"
     protein_bind    complement(3198..3297)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3314..3333)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3315..3333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3338..3354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3338..3354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3456..3475)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3467..4276
                     /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(3534..3553)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4423..5011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4912..4931
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctac aaaatgggta agacacaaaa gaagaatagt aagggacgtt tagatagata
      721 ctattatcta gcaaaggaga aaggttatcg tgctcgttca tcctttaaga ttattcagat
      781 taatgaaaag tatggccact tcttagaaaa atcgaaggtt gttattgatc tgtgtgctgc
      841 tcctggttca tggtgtcaag ttgcatccaa actctgtcct gtcaactcct taattattgg
      901 tgttgatatt gttccaatga agccgatgcc caatgttata acttttcaaa gtgatattac
      961 aacagaagat tgtagatcaa aattgagggg ttatatgaag acttggaaag ctgatacagt
     1021 gttgcatgat ggtgctccta atgttggttt gggttgggtt caggatgctt tcacccaatc
     1081 tcaattaacc ttacaagctt tgaagttggc cgtcgaaaat ttggtggtga acggtacttt
     1141 tgttactaaa attttcagat ccaaggatta taataaattg atttgggttt tccaacaatt
     1201 gtttgagaaa gttgaagcca caaaaccacc cgcatcaaga aatgtttctg cagaaatttt
     1261 tgtagtgtgt aaaggtttca aggcaccaaa gaggctggac ccaagattac tggatccaaa
     1321 agaagttttt gaagaattgc cagatgggca acagaatatg gagtccaaga tttataatcc
     1381 cgaaaaaaaa gttagaaaaa gacaaggtta tgaggaaggt gataatttac tgtatcatga
     1441 aacctcaatt ttggattttg tcaggacgga agacccaata agcatgcttg gagaaatgaa
     1501 taagtttaca attgatgaga atgaccatga atggaagatc ttgaagaaat tgaaacaaac
     1561 cacagacgag ttccgttcct gtattgaaga tttaaaagtc ttgggtaaaa aggattttaa
     1621 aatgatccta agatggagaa agattgcaag ggaaatcttg ggcatcgaag ttaaggacga
     1681 cgctaaaaca gaaattgaag tagtaccgtt aacagaagag gagcaaattg aaaaagattt
     1741 gcaaggttta caggagaagc agcgtctaaa tgtcaagcgc gaaagaagaa ggaagaacga
     1801 aatgaagcaa aaggaactac agagaatgca aatgaacatg ataaccccca ctgatattgg
     1861 tattgaagcc gcaagtttgg gtaaagaatc gttgtttaat ttgaaaactg cagaaaagac
     1921 tggtatcttg aacgacttgg cgaagggtaa gaaaagaatg atttttacag acgatgaatt
     1981 agccaaagat aatgatattt acattgatga aaacatcatg atcaaagata aggattctgc
     2041 cgctgatgcg gacgatttag aaagcgagtt gaatgccatg tatagtgatt ataaaactag
     2101 aaggtcggaa agggatgcca agtttagagc taagcaagca cgtggtggcg ataatgagga
     2161 agaatggacc ggttttaatg agggaagtct agaaaagaaa gaagaggaag gtaaagatta
     2221 tatcgaagac aatgacgatg aaggtgtcga aggagactcc gatgatgatg aagccatcac
     2281 caatttgatt agcaaattga agggacaaga aggtgatcac aaattgagta gtaaggcacg
     2341 tatgattttc aatgatccga tatttaataa cgttgaacct gatttgccag taaataccgt
     2401 caatgatggc ataatgagtt ccgagtctgt tggtgatatt tctaaattaa ataagaaaag
     2461 aaaacatgaa gaaatgcacc agaaacaaga tgaagcagat tcttcggatg agagttcaag
     2521 tgatgactcc gatttcgaaa ttgtggccaa cgacaatgca tcagaagaat tcgattctga
     2581 ttatgattca gaggaagaaa aaaatcaaac aaagaaagaa aagcattcca gagacattga
     2641 tattgctact gttgaagcca tgactttggc acatcagtta gcattgggtc agaaaaacaa
     2701 gcatgatctt gttgatgaag gtttcaatag atacaccttc cgtgacaccg aaaatttgcc
     2761 agattggttt ttagaagatg aaaaggaaca ctccaagata aataagccga tcactaagga
     2821 ggcagcgatg gcaattaaag aaaaaataaa ggcaatgaac gcccgtccta ttaagaaggt
     2881 tgctgaagct aaggcaagaa agaggatgcg tgccgtggct cggttagaaa agatcaagaa
     2941 gaaggcgggc ttgattaacg atgattcaga caagacagag aaggacaagg ctgaagaaat
     3001 ttctagattg atgcgtaagg ttaccaaaaa accaaagacc aagccaaagg ttactttggt
     3061 tgttgcctcg ggtaggaaca aaggtttggc aggtagacca aagggcgtta agggtaagta
     3121 taagatggtt gatggtgtca tgaagaatga acaaagagcc ctaagacgta tcgcaaagaa
     3181 gcatcacaag aaaaagaacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     3241 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3301 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     3361 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaataa tatcatcatg
     3421 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     3481 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     3541 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     3601 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     3661 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     3721 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     3781 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3841 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3901 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3961 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     4021 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     4081 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     4141 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     4201 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     4261 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     4321 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     4381 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4441 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4501 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4561 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4621 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4681 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4741 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4801 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4861 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4921 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4981 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5041 cctggccttt tgctggcctt ttgctcacat gtt
//
