LOCUS       sequence-191954-        3666 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Shuttle yeast Cka1 into Gateway destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3666)
  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 3666)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Cka1
FEATURES             Location/Qualifiers
     source          1..3666
                     /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(1255..1354)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1371..1390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1372..1390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1395..1411)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1395..1411)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1513..1532)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1524..2333
                     /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(1591..1610)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2480..3068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2969..2988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3222..3239
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3398..3425)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3517..3603)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3652..3666,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3666,1..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 aaatgcaggg tatggtcaga ggctcgtgtt tatacgaata
      181 tcaataaaca aagaaccgag gaatattggg attatgaaaa tactgtaatt gattggtcca
      241 caaatacaaa ggactatgaa attgaaaata aagttgggcg aggaaaatac tccgaggttt
      301 tccaaggtgt caaattagac tctaaagtta aaattgttat taagatgttg aaaccagtta
      361 aaaagaagaa gatcaagaga gaaatcaaaa tcttaacgga tttgtctaac gaaaaagtgc
      421 ctccaacgac tttgccattt caaaaagatc aatattacac aaatcaaaag gaggacgttt
      481 taaaattcat caggccctat atctttgatc agccacacaa tggtcatgca aacataattc
      541 atctatttga tataataaag gatcccatct caaaaactcc ggctttggtc ttcgaatacg
      601 tagataatgt ggacttccgt attctttacc ctaaattaac cgatctcgaa attaggtttt
      661 atatgtttga gttattaaag gccttagact attgtcattc aatgggaata atgcatagag
      721 atgttaaacc tcataacgta atgattgatc ataagaataa aaaattgcga ttgatagatt
      781 gggggcttgc tgaattttat catgttaata tggaatacaa tgttcgtgtt gcgtcgaggt
      841 tttttaaggg tcctgaacta ctagttgact acagaatgta tgattattct ttagacttgt
      901 ggtcgtttgg gacaatgttg gcttctatga tctttaaaag agagccattt ttccatggaa
      961 cgagtaacac agaccagctt gtcaagatcg tcaaagtact tggtacaagc gattttgaga
     1021 aatacctgtt gaagtatgaa attaccttac cgagagaatt ttacgatatg gaccaataca
     1081 tcagaaagcc ttggcataga ttcatcaatg atggtaataa acatttaagc ggcaacgatg
     1141 aaattattga ccttattgac aatcttttga gatatgatca tcaagaaaga ttaactgcta
     1201 aggaagcgat gggacacccg tggtttgccc caataaggga acaaattgaa aaaaacccag
     1261 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     1321 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     1381 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     1441 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1501 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1561 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1621 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1681 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1741 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1801 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     1861 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1921 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     1981 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2041 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2101 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2161 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2221 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2281 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     2341 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     2401 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     2461 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2521 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2581 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2641 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2701 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2761 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2821 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2881 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2941 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3001 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3061 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3121 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3181 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3241 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3301 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
     3361 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
     3421 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     3481 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     3541 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     3601 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
     3661 gacgtt
//
