LOCUS       sequence-191956-        3566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle yeast Cka2 into Gateway destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3566)
  AUTHORS   Vincent NG, Charette JM, Baserga SJ
  TITLE     The SSU processome interactome in Saccharomyces cerevisiae reveals 
            potential new protein subcomplexes.
  JOURNAL   RNA. 2017 Oct 20. pii: rna.062927.117. doi: 10.1261/rna.062927.117.
  PUBMED    29054886
REFERENCE   2  (bases 1 to 3566)
  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"; journalName: "RNA. 2017 
            Oct 20. pii: rna.062927.117. doi: 10.1261/rna.062927.117."
COMMENT     Sequence Label: pDONR221-Cka2
FEATURES             Location/Qualifiers
     source          1..3566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1155..1254)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1271..1290)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1272..1290)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1295..1311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1295..1311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1413..1432)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1424..2233
                     /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(1491..1510)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2380..2968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2869..2888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3122..3139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3298..3325)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3417..3503)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3552..3566,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3566,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctacaaaatg ccattacctc cgtcaacatt gaaccagaaa tctaatagag
      181 tctactctgt agctagggtg tacaagaatg cctgcgagga gagaccacaa gaatactggg
      241 actacgaaca aggggtgacc atcgattggg gaaagatttc caattacgaa attatcaaca
      301 aaattggaag agggaaatat tccgaagtgt tcagcggtag atgtattgta aacaaccaga
      361 agtgtgttat taaagtttta aaaccagtta aaatgaaaaa aatttataga gagttgaaaa
      421 ttctgaccaa tctaacaggc ggccccaatg ttgttggcct ttatgatata gtacaagacg
      481 ctgactccaa aatacctgct ttgatctttg aggaaatcaa aaatgttgat ttcagaactt
      541 tatatcctac attcaaactt cctgacatcc agtattattt cacgcaatta ttgattgcgt
      601 tagactactg tcactccatg ggcataatgc acagagacgt aaagcctcag aatgtcatga
      661 ttgatcctac ggaacgtaaa ctaaggctga tcgattgggg cctggcggag ttctaccatc
      721 caggtgtaga ttacaacgtt cgtgtcgctt cgcgttacca caagggacca gaacttttag
      781 taaacttgaa ccaatatgac tactccctag acttatggtc agtaggatgc atgctagcag
      841 ctattgtctt caaaaaagaa ccttttttca aagggtcgtc taatccagat caactggtaa
      901 agattgccac agtactagga accaaggaac tgttaggcta tttgggtaag tacgggttgc
      961 acttaccatc tgaatacgac aacattatga gagactttac aaaaaaatcg tggacacact
     1021 ttataacctc cgagaccaaa ttagctgttc ctgaagtggt tgatttaatc gacaatttat
     1081 taaggtatga ccatcaagaa agattaacag caaaggaggc tatggatcat aagtttttca
     1141 aaacgaagtt tgaaacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1201 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1261 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1321 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1381 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1441 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1501 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     1561 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1621 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1681 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     1741 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1801 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1861 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     1921 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     1981 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2041 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2101 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2161 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2221 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2281 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2341 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2401 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2461 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2521 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2581 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2641 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2701 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2761 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2821 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2881 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2941 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3001 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3061 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3121 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3181 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3241 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     3301 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     3361 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     3421 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     3481 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     3541 atggatgttt tcccagtcac gacgtt
//
