LOCUS       sequence-338615-        5302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 2 with attR4 and R3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5302)
  AUTHORS   Nakazato I, Okuno M, Yamamoto H, Tamura Y, Itoh T, Shikanai T, 
            Takanashi H, Tsutsumi N, Arimura SI
  TITLE     Targeted base editing in the plastid genome of Arabidopsis thaliana.
  JOURNAL   Nat Plants. 2021 Jul;7(7):906-913. doi: 10.1038/s41477-021-00954-6. 
            Epub 2021 Jul 1.
  PUBMED    34211131
REFERENCE   2  (bases 1 to 5302)
  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/s41477-021-00954-6"; journalName: "Nat Plants"; date: 
            "2021-07"; volume: "7"; issue: "7"; pages: "906-913"
COMMENT     Sequence Label: E2_pENTR_R4R3_T CTP
FEATURES             Location/Qualifiers
     source          1..5302
                     /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    3365..3488
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3543..3562)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3544..3562)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3567..3583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3567..3583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3685..3704)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3696..4505
                     /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(3763..3782)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4652..5240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5141..5160
                     /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 actctagcgt 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 catggattca cagctagtct tgtctctgaa gctgaatcca
     3241 agcttcactc ctctttctcc tctcttccct ttcactccat gttcttcttt ttcgccgtcg
     3301 ctccggtttt cttcttgcta ctcccgccgc ctctattctc cggttaccgt ctacgccgcg
     3361 aagacaactt tgtataataa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     3421 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     3481 tcactatgaa tcaactactt agatggtatt agtgacctgt actgcagggc ggccgcgata
     3541 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     3601 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     3661 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     3721 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     3781 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     3841 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     3901 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     3961 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     4021 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     4081 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     4141 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     4201 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     4261 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     4321 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     4381 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     4441 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     4501 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     4561 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     4621 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4681 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4741 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4801 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4861 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4981 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5041 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5101 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5161 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5221 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5281 gctggccttt tgctcacatg tt
//
