LOCUS       sequence-305029-        2644 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 2644)
  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 2644)
  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: pENTR2-R2-PBR-L4
FEATURES             Location/Qualifiers
     source          1..2644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(108..231)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   506..540
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    complement(545..640)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(753..772)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             764..1573
                     /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(831..850)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1716..2304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2205..2224
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2439..2466)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2558..2644)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atgcctggca gttccctact ctcgcgtcaa cgctagcatg gatctcgggc cctgcagctc
       61 tagagctcga attctacagg tcactaatac catctaagta gttggttcat agtgactgca
      121 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
      181 tgatatttat atcattttac gtttctcgtt caactttctt gtacaaagtg ggatatctat
      241 aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa
      301 ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc
      361 acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg
      421 gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga
      481 aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg
      541 ttaacaactt ttctatacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
      601 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgcagctctg
      661 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
      721 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
      781 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
      841 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
      901 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
      961 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1021 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     1081 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     1141 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     1201 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     1261 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     1321 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     1381 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     1441 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     1501 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     1561 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     1621 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttttc gttccactga
     1681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1861 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1921 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1981 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2041 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2101 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2161 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2221 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2401 cgtattaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     2461 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     2521 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     2581 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     2641 tttg
//
