LOCUS       sequence-311216-        4107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant mitoTALEN vector assembly, Entry clone 2 with attL1 L4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4107)
  AUTHORS   Kazama T, Okuno M, Watari Y, Yanase S, Koizuka C, Tsuruta Y, Sugaya 
            H, Toyoda A, Itoh T, Tsutsumi N, Toriyama K, Koizuka N, Arimura SI
  TITLE     Curing cytoplasmic male sterility via TALEN-mediated mitochondrial 
            genome editing.
  JOURNAL   Nat Plants. 2019 Jul;5(7):722-730. doi: 10.1038/s41477-019-0459-z. 
            Epub 2019 Jul 8.
  PUBMED    31285556
REFERENCE   2  (bases 1 to 4107)
  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"; doi: 
            "10.1038/s41477-019-0459-z"; journalName: "Nat Plants"; date: 
            "2019-07"; volume: "5"; issue: "7"; pages: "722-730"
COMMENT     Sequence Label: pENTR E2 HSPter-35Sp-mito
FEATURES             Location/Qualifiers
     source          1..4107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..346
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     289..309
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    366..421
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     protein_bind    607..730
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(785..804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(786..804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(809..825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(809..825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(927..946)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             938..1747
                     /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(1005..1024)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1894..2482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2383..2402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2636..2653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2812..2839)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2931..3017)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3066..3088
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3080..3097
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3081..3097
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3147..3270)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3275..3523
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
ORIGIN
        1 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
      121 cgataaagga aaggctatcg ttcaagatgc ctctaccgac agtggtccca aagatggacc
      181 cccacccacg aggaacatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
      241 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag aggacaggct tcttgagatc
      361 cttcaacaat taccaacaac aacaaacaac aaacaacatt acaattacta tttacaatta
      421 cagtcgaccc acgcgtccga taatttcccc aatttctcag agagactcaa aatcaccacc
      481 accaccaaat cgccgccatg tttaaacaag cttctcgtct cctctcccga tctgtcgccg
      541 ccgcatcttc caaatcggtg acgactcgtg ccttttcaac ggaacttcca tcgacgctcg
      601 attccacaac tttgtataat aaagttgaac gagaaacgta aaatgatata aatatcaata
      661 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
      721 agtcactatg aatcaactac ttagatggta ttagtgacct gtactgcagg gcggccgcga
      781 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
      841 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
      901 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
      961 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1021 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1081 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1141 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1201 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1261 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1321 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1381 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     1441 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     1501 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1561 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1621 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1681 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1741 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     1801 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     1861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2041 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2581 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2641 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2701 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2761 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     2821 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     2881 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     2941 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     3001 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     3061 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc ccctacaggt
     3121 cactaatacc atctaagtag ttgattcata gtgactggat atgttgtgtt ttacagtatt
     3181 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     3241 tttctcgttc aacttttcta tacaaagttg tgaatatgaa gatgaagatg aaatatttgg
     3301 tgtgtcaaat aaaaagcttg tgtgcttaag tttgtgtttt tttcttggct tgttgtgtta
     3361 tgaatttgtg gctttttcta atattaaatg aatgtaagat ctcattataa tgaataaaca
     3421 aatgtttcta taatccattg tgaatgtttt gttggatctc ttctgcagca tataactact
     3481 gtatgtgcta tggtatggac tatggaatat gattaaagat aagatgggct catagagtaa
     3541 aacgaggcga gggacctata aacctccctt catcatgcta tttcatgatc tattttataa
     3601 aataaagatg tagaaaaaag taagcgtaat aaccgcaaaa caaatgattt aaaacatggc
     3661 acataatgag gagattaagt tcggtttacg tttattttag tactaattgt aacgtgagac
     3721 tacgtatcgg gaatcgccta attaaagcat taatgcgaac ctgattagat tcaccgaccc
     3781 tcctatcgtg tcgacctttc tgtttcttag aattttttgg tagtctatgt actaataatg
     3841 tcagcttcgt atttatttca taagcaattt gcatttgcaa tttgtttttt acttttattt
     3901 ttattgtatt gtggaatgtg gactcgtacc aacatgaagt tatataccac caaaaaaatt
     3961 acagttagtc aaaagattca cgagtgagag ctacttatga ttgtctttta cgtatatgtc
     4021 taattgtcta tttgctcaat aatctttgta ctttcttttg tcgttgataa atctagagga
     4081 tcccccctca gaagaccaga gggctat
//
