LOCUS       sequence-301959-        2342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutisite Gateway mediated vector construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2342)
  AUTHORS   Yoshimatsu S, Sone T, Nakajima M, Sato T, Okochi R, Ishikawa M, 
            Nakamura M, Sasaki E, Shiozawa S, Okano H
  TITLE     A versatile toolbox for knock-in gene targeting based on the 
            Multisite Gateway technology.
  JOURNAL   PLoS One. 2019 Aug 27;14(8):e0221164. doi: 
            10.1371/journal.pone.0221164. eCollection 2019.
  PUBMED    31454364
REFERENCE   2  (bases 1 to 2342)
  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: "PLoS One.";
            date: "2019-08-27"; volume: "14(8):e0221164. doi"; pages: " 
            10.1371/journal.pone.0221164. eCollection 2019"
COMMENT     Sequence Label: pENTR-R2L4
FEATURES             Location/Qualifiers
     source          1..2342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(202..221)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             213..1022
                     /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(280..299)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1165..1753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1654..1673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1888..1915)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2007..2093)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    complement(2201..2325)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(join(2336..2342,1..89))
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tctatacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
       61 cactatcagt caaaataaaa tcattatttg ccatccagct gcagctctgg cccgtgtctc
      121 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
      181 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
      241 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
      301 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
      361 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
      421 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
      481 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
      541 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
      601 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
      661 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
      721 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
      781 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
      841 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
      901 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
      961 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     1021 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     1081 gcgcaagctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1861 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     1921 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     1981 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     2041 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
     2101 gcagttccct actctcgcgt taacgctagc atggatctcg ggccctgcag ctctagagct
     2161 cgaattctac aggtcactaa taccatctaa gtagttggtt catagtgact gcatatgttg
     2221 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     2281 tatatcattt tacgtttctc gttcaacttt cttgtacaaa gtggtctcga gttaacaact
     2341 tt
//
