LOCUS       sequence-379070-        4225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 3 with attL2 and L3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4225)
  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 4225)
  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: E3_pENTR_L3-L2_NI_G1333-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(919..1728)
                     /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     1642..1661
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1720..1739
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1841..1857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1862..1880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1911..2010
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2032..2283)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    2805..2826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2841..2846,2847..2864,2865..2871)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2879..2895
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2884..2906
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2903..2919
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2915..3325
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     2938..2958
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2940..2958
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3006..3024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3034..3050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3635..3679)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    complement(3682..3777)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3794..3810)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3874..3960
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4052..4079
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
      841 ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa
      901 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      961 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1021 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1081 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1141 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1201 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1261 gttattcatt cgtgattgcg cctgagcaag gcgaaatacg cgatcgctgt taaaaggaca
     1321 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1381 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt
     1441 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1501 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1561 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     1621 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1681 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt
     1741 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     1801 aatgtaacat cagagatttt gagacacggg ccagagctgc caggaaacag ctatgaccat
     1861 gtaatacgac tcactatagg ggatatcgcg gccgccctgc agctggatgg caaataatga
     1921 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttctta
     1981 taatgccaac tttgtacaag aaagctgggt tctagaggat cgctcgagtt atcacaacat
     2041 cttgatcttg ttctctccat tggaatcctg gataactaac gcccaaggtt tgtactcagg
     2101 agcatctgag gttagaagca ttacgttctc atccgtcgat tcgtcatagg cagtatgaac
     2161 aagtatatcg ctttcgggct tattgcctat cacttcctca acctcctctg gtaacatcag
     2221 gattgactcc tgaatgacta gttgctttcc agtctctttc tcaatgatgt cagagagatt
     2281 tgtagatcct ccactacacc cacctttcgt tgggcttttg ggtgaattgg aattcccagt
     2341 aaacaccttg gtttcaccag tagcacctct tttgacagga atagcacctt caggtggaac
     2401 tactgtcatc ttagcgtttt ctgggagaag ggtttctgtc atgttgacgc aaaatccaca
     2461 tgttccttca gggttgttgt ggaacaccaa cccttcagaa atgccgttat ccctcatgaa
     2521 caatgcagat tgaccttcaa catgtcctgc attggcgtaa ttcggatacg gagttggact
     2581 accccccaat ccctttttca ctgcatccag cgcaggacgc ccgccgaggc aggccaaggc
     2641 gacgaggtgg tcgttggtca acgcggccaa cgccggatca gggcgagata actgggcaac
     2701 aattgactcc agtgcgggtc tgccccctat gttggatgca atagccacta cctgctcagg
     2761 ggtggagacg accggtggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2821 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2881 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     2941 attaaccctc actaaaggga acaaaagctg gaggacctcg agggggggcc cggtacccaa
     3001 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     3061 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3121 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3181 tggcgaatgg gacgcgccct gtagcggcgc ccgtctcctt gagcggggca cccgtcagtg
     3241 cattgcgcca tgcatgcact gcctccactg cggtcacgcc gccacgtttt gcaatcttga
     3301 gaagttggcc tgtgtccaac tgtaacggtg gacctctcaa ctctcccgcc accgtgagca
     3361 aggcctccag agcgcgtgcg ccggaccact gtttgccgac gccaacgatc gcttcgtgtg
     3421 tcgcctctgg caacgctgcg atcatgtcct gatacttgac agcgacggtc cctaacgctg
     3481 ccgggtgttg gcttaacgca acgatgtgcg cgtgtgtaaa cccgtggccg accagtgcct
     3541 cgtggtgctg cgccactgtc gaacgaacct tcggtttgat cttctcctgt tgctgctggc
     3601 tgtagccgag cgtgcgtaga tcggcgatat cgatgctgtc catgtgctgt ctctcgaact
     3661 tggcagcggc ggtctcctta tcaactttat tatacaaagt tggcattata aaaaagcatt
     3721 gctcatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttggg
     3781 gcccgagctt aagactggcc gtcgttttac aacgtcgtga ctgggaaaac atccatgcta
     3841 gcgttaacgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
     3901 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3961 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
     4021 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
     4081 gcgtttctac aaactcttcc tggctagcgg tacgcgtatt aattgcgttg cgctcactgc
     4141 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4201 ggagaggcgg tttgcgtatt gggcg
//
