LOCUS       sequence-302007-        2907 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 2907)
  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 2907)
  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-L5-CMV-R3
FEATURES             Location/Qualifiers
     source          1..2907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    29..124
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        131..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    731..854
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(991..1010)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1002..1811
                     /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"
     rep_origin      1954..2542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2443..2462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2677..2704)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2796..2882)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttaacgcta gcatggatct cgggccccaa ataatgattt tattttgact gatagtgacc
       61 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtatacaaaa
      121 gttgaagctt cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccgaagctt caactttgta taataaagtt gaacgagaaa cgtaaaatga tataaatatc
      781 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
      841 atccagtcac tatgaatcaa ctacttagat ggtattagtg acctgtactg cagctctggc
      901 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
      961 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     1021 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     1081 ctcgtgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     1141 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     1201 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     1261 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     1321 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     1381 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     1441 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     1501 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     1561 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     1621 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     1681 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     1741 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     1801 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     1861 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2641 tattaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     2701 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     2761 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     2821 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     2881 tgatgcctgg cagttcccta ctctcgc
//
