LOCUS       sequence-192203-        4851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle yeast Pfa1 into Gateway destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4851)
  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 4851)
  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-Pfa1
FEATURES             Location/Qualifiers
     source          1..4851
                     /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(2440..2539)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2556..2575)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2557..2575)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2580..2596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2580..2596)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2698..2717)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2709..3518
                     /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(2776..2795)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3665..4253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4154..4173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4407..4424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4583..4610)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4702..4788)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4837..4851,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4851,1..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 gcaaaaaggc atagtcatta tcaaggaagc agacgtaggc
      181 atgcacgtgg tagcaattcc aagaaagctg gcagggggaa tgctaaaggc atacaaggca
      241 gaaagataaa gaaaaaacct actcccacta acagttggca taactcttct ataccactag
      301 gagaaggtga tcttgatgat gtaggcgccg acttcaaccc aggaagagca ttcatatcac
      361 caaaaactat agaagactat tattttggtc gcgatgccaa gagccgttct atgaaaatgg
      421 gtggtttaag accaggtaat cgatatgatt cttcaactga cctacaagca ggaagggcag
      481 cattccgcaa aagacctatg caatttgtaa aagctaagga agtttacgat ccatcccaca
      541 atatgattca aaaactgaga gcaaagaacg aaacaaagaa cagtgaagaa attgtcgaaa
      601 gggaagcaga cgtttttgaa gaaccaggaa agatgacttc tgatgttgaa tatattaaca
      661 atgaagacag tgaaaacgag gatgatgata gtcagaattc accctcgacg gaccactcgc
      721 tgtcttcgaa tgaaagcaaa gttgaagatg gtgacctgtt ttttgtagat gaagaggctc
      781 agcaatcgcc ggatctaacc aaaataaaaa gagtatgtat cgaggaaatc gcaaggccaa
      841 gagaagttgc gatagagttc gatcctatct taacaattgg aaaggttgaa cttagcgttt
      901 ccgaaggtaa cgaaagcaag gaaattagtg ttgacgttcc aaataagggt aataagacat
      961 atcatccatt tgctggttac atttccaacg tattacatgg tatgcatact agcgactcgg
     1021 acaatgatga actagactac gaaatagaaa cagagaataa tagtgaacct ttatacgaat
     1081 cctcagcaag ttctgaggtt gatcaagggt ttaattatgt tggccaacgt cacaattccc
     1141 gagctgacaa caatctttta ccaagcccta gtccgcagtt gacagaagat ataaaatgtc
     1201 tttcaataaa tggtaccaaa acattcgaag gcaataatga taatctgcca tctcctgcta
     1261 gcgaagagtt agaatttgga ttcaaagaag aagattttgt cataaatact aatgacatag
     1321 tggtctccaa tataagaatg ggcggagtag acaattcata ttatctacgg tgttacagat
     1381 tactgggaga ttacgacttt cattggattg accaggactt actcacagac ttcgttgtag
     1441 acgagttagg tcttcctgaa gatagattac ctgcatattt gaattttatt aaaaattctc
     1501 ttataccaaa aattgaacca gctgagccaa cctactcaga tattccgatt tctgattcga
     1561 gtgatgaagg tgacagctat gaaggcgaca gctatgaaga tgatgaagat atggcctcct
     1621 cggttgtgca tagcgacatt gaagagggtc tggacgacct aattgcatac acattgaaac
     1681 atgatacaga aagattcaaa acatttgaaa ccaagtcctt ggaaactaag ggtaaaggta
     1741 agaagaagaa gctactcatc gatgacgcct tagcgctgga tacagaaacg ttggagacgt
     1801 tacaaagtaa gtttagtaaa cgcatagaaa ccaaagcaaa gaaaagaaaa gccaaagaag
     1861 atttcattga tcaggagaat agaaattcga acgatatgct caagaaatac ccatatggac
     1921 tccatattca aaatattaaa gatgaatttg agtccttttt gtctaggaac aatgatagac
     1981 tgactttccc tccattagat cctcatggaa acaagaccgt aatgaaaatt gcgaagcatt
     2041 ataatatgaa gagttcaaaa attgggaaag caaatcatac gtcggttgtt gtagaaaaaa
     2101 taaagaaaac gaaatggtct tctccgaatt atagtctcat tgaccaattg atgaggcaga
     2161 ggccagtatt catgagaata gatataagaa gaccgaggga ggaacaagct gctttcgaaa
     2221 gaactaaaac aatcagaggg aaatttcatg ttaaagaagg tgagattgtt ggtcaaaacg
     2281 ctccagagat tggaaacgaa aatatcggta gaagaatgtt ggaaaagctt gggtggaaaa
     2341 gtggagaggg ccttggtatt caaggcaata aaggtataag tgaaccaatt tttgccaaaa
     2401 ttaaaaagaa tagatcgggt ttaagacaca gtgaaagtaa cccagctttc ttgtacaaag
     2461 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     2521 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     2581 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2641 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2701 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2761 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2821 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2881 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2941 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     3001 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     3061 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     3121 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     3181 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     3241 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     3301 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     3361 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     3421 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     3481 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     3541 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3601 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3661 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3721 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3781 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3841 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3901 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3961 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4021 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4081 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4141 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4201 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4261 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4321 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4381 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4441 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4501 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     4561 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     4621 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4681 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4741 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     4801 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt t
//
