LOCUS       sequence-381638-        5164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nTALEN or nTALECD vector assembly, Entry clone for plants to express
            ORFs from entry clone with NLS under control of the RPS5A promoter 
            and HSP terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5164)
  AUTHORS   Hosoda A, Nakazato I, Takanashi H, Tsutsumi N, Arimura SI
  TITLE     TALE-based C-to-T base editor for multiple homologous genes with 
            flexible precision
  JOURNAL   bioRxiv 2023.02.27.530312
REFERENCE   2  (bases 1 to 5164)
  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: "bioRxiv 
            2023.02.27.530312"
COMMENT     Sequence Label: E2_pENTR_R4-R3_HSPter-RPS5A-NLS
FEATURES             Location/Qualifiers
     source          1..5164
                     /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)"
     CDS             3205..3225
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    3227..3350
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3405..3424)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3406..3424)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3429..3445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3429..3445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3547..3566)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3558..4367
                     /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(3625..3644)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4514..5102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5003..5022
                     /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 catgcctaag aagaagagga aggttacaac tttgtataat
     3241 aaagttgaac gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat
     3301 aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg aatcaactac
     3361 ttagatggta ttagtgacct gtactgcagg gcggccgcga tatcccctat agtgagtcgt
     3421 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     3481 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     3541 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     3601 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     3661 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     3721 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     3781 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     3841 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     3901 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     3961 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     4021 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     4081 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     4141 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     4201 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     4261 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     4321 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     4381 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     4441 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     4501 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4561 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4621 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4681 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4741 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4801 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4861 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4921 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4981 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5041 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5101 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5161 tgtt
//
