LOCUS       sequence-316149-        5248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant mitoTALEN vector assembly, Entry clone 2 with attL1 L4; 
            Multisite Gateway entry vector to terminate the E1 ORF and having 
            RPS5A promoter with mitochondrial presequence for the E3 ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5248)
  AUTHORS   Arimura SI, Ayabe H, Sugaya H, Okuno M, Tamura Y, Tsuruta Y, Watari 
            Y, Yanase S, Yamauchi T, Itoh T, Toyoda A, Takanashi H, Tsutsumi N
  TITLE     Targeted gene disruption of ATP synthases 6-1 and 6-2 in the 
            mitochondrial genome of Arabidopsis thaliana by mitoTALENs.
  JOURNAL   Plant J. 2020 Oct 24. doi: 10.1111/tpj.15041.
  PUBMED    33098708
REFERENCE   2  (bases 1 to 5248)
  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: "Plant J. 
            2020 Oct 24. doi: 10.1111/tpj.15041."
COMMENT     Sequence Label: pENTR E2 HSPter-RPS5Ap-mito
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..726)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      731..979
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    3311..3434
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3489..3508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3490..3508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3513..3529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3513..3529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3631..3650)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3642..4451
                     /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(3709..3728)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4598..5186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5087..5106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgtgaa tatgaagatg aagatgaaat atttggtgtg tcaaataaaa agcttgtgtg
      781 cttaagtttg tgtttttttc ttggcttgtt gtgttatgaa tttgtggctt tttctaatat
      841 taaatgaatg taagatctca ttataatgaa taaacaaatg tttctataat ccattgtgaa
      901 tgttttgttg gatctcttct gcagcatata actactgtat gtgctatggt atggactatg
      961 gaatatgatt aaagataaga tgggctcata gagtaaaacg aggcgaggga cctataaacc
     1021 tcccttcatc atgctatttc atgatctatt ttataaaata aagatgtaga aaaaagtaag
     1081 cgtaataacc gcaaaacaaa tgatttaaaa catggcacat aatgaggaga ttaagttcgg
     1141 tttacgttta ttttagtact aattgtaacg tgagactacg tatcgggaat cgcctaatta
     1201 aagcattaat gcgaacctga ttagattcac cgaccctcct atcgtgtcga cctttctgtt
     1261 tcttagaatt ttttggtagt ctatgtacta ataatgtcag cttcgtattt atttcataag
     1321 caatttgcat ttgcaatttg ttttttactt ttatttttat tgtattgtgg aatgtggact
     1381 cgtaccaaca tgaagttata taccaccaaa aaaattacag ttagtcaaaa gattcacgag
     1441 tgagagctac ttatgattgt cttttacgta tatgtctaat tgtctatttg ctcaataatc
     1501 tttgtacttt cttttgtcgt tgataaatct agaggatccc cctcaacttt tgattcgcta
     1561 tttgcagtgc acctgtggcg ttcatcacat cttttgtgac actgtttgca ctggtcattg
     1621 ctattacaaa ggaccttcct gatgttgaag gagatcgaaa gtaagtaact gcacgcataa
     1681 ccattttctt tccgctcttt ggctcaatcc atttgacagt caaagacaat gtttaaccag
     1741 ctccgtttga tatattgtct ttatgtgttt gttcaagcat gtttagttaa tcatgccttt
     1801 gattgatctt gaataggttc caaatatcaa ccctggcaac aaaacttgga gtgagaaaca
     1861 ttgcattcct cggttctgga cttctgctag taaattatgt ttcagccata tcactagctt
     1921 tctacatgcc tcaggtgaat tcatctattt ccgtcttaac tatttcggtt aattaaagca
     1981 cgaacaccat tactgcatgt agaagcttga taaactatcg ccaccaattt atttttgttg
     2041 cgatattgtt actttcctca gtatgcagct ttgaaaagac caaccctctt atcctttaac
     2101 aatgaacagg tttttagagg tagcttgatg attcctgcac atgtgatctt ggcttcaggc
     2161 ttaattttcc aggtaaagca ttatgagata ctcttatatc tcttacatac ttttgagata
     2221 atgcacaaga acttcataac tatatgcttt agtttctgca tttgacactg ccaaattcat
     2281 taatctctaa tatctttgtt gttgatcttt ggtagacatg ggtactagaa aaagcaaact
     2341 acaccaaggt aaaatacttt tgtacaaaca taaactcgtt atcacggaac atcaatggag
     2401 tgtatatcta acggagtgta gaaacatttg attattgcag gaagctatct caggatatta
     2461 tcggtttata tggaatctct tctacgcaga gtatctgtta ttccccttcc tctagctttc
     2521 aatttcatgg tgaggatatg cagttttctt tgtatatcat tcttcttctt ctttgtagct
     2581 tggagtcaaa atcggttcct tcatgtacat acatcaagga tatgtccttc tgaattttta
     2641 tatcttgcaa taaaaatgct tgtaccaatt gaaacaccag ctttttgagt tctatgatca
     2701 ctgacttggt tctaaccaaa aaaaaaaaaa tgtttaattt acatatctaa aagtaggttt
     2761 agggaaacct aaacagtaaa atatttgtat attattcgaa tttcactcat cataaaaact
     2821 taaattgcac cataaaattt tgttttacta ttaatgatgt aatttgtgta acttaagata
     2881 aaaataatat tccgtaagtt aaccggctaa aaccacgtat aaaccaggga acctgttaaa
     2941 ccggttcttt actggataaa gaaatgaaag cccatgtaga cagctccatt agagcccaaa
     3001 ccctaaattt ctcatctata taaaaggagt gacattaggg tttttgttcg tcctcttaaa
     3061 gcttctcgtt ttctctgccg tctctctcat tcgcgcgacg caaacgatct tcaggtgatc
     3121 ttctttctcc aaatcctctc tcataactct gatttcgtac ttgtgtattt gagctcacgc
     3181 tctgtttctc tcaccacagc catgtttaaa caagcttctc gtctcctctc ccgatctgtc
     3241 gccgccgcat cttccaaatc ggtgacgact cgtgcctttt caacggaact tccatcgacg
     3301 ctcgattcca caactttgta taataaagtt gaacgagaaa cgtaaaatga tataaatatc
     3361 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     3421 atccagtcac tatgaatcaa ctacttagat ggtattagtg acctgtactg cagggcggcc
     3481 gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     3541 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     3601 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     3661 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     3721 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     3781 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     3841 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     3901 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     3961 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4021 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     4081 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4141 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     4201 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4261 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4321 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4381 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4441 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     4501 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     4561 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4621 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4681 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4741 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4801 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4861 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4921 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4981 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5041 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5101 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5161 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5221 ccttttgctg gccttttgct cacatgtt
//
