LOCUS       sequence-379217-        4267 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 4267)
  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 4267)
  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_HD_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..126
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(436..846)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     711..727
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        737..755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(803..823)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(803..821)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(842..858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(855..877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    866..882
                     /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)."
     promoter        complement(join(890..896,897..914,915..920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    935..956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1520..1771
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    complement(1793..1892)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1923..1941)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1946..1962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2064..2083)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2075..2884
                     /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(2142..2161)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3031..3619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3520..3539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3773..3790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3949..3976)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4068..4154)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4218..4234
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(4251..4267,1..79)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
       61 actttgtata ataaagttga taaggagacc gccgctgcca agttcgagag acagcacatg
      121 gacagcatcg atatcgccga tctacgcacg ctcggctaca gccagcagca acaggagaag
      181 atcaaaccga aggttcgttc gacagtggcg cagcaccacg aggcactggt cggccacggg
      241 tttacacacg cgcacatcgt tgcgttaagc caacacccgg cagcgttagg gaccgtcgct
      301 gtcaagtatc aggacatgat cgcagcgttg ccagaggcga cacacgaagc gatcgttggc
      361 gtcggcaaac agtggtccgg cgcacgcgct ctggaggcct tgctcacggt ggcgggagag
      421 ttgagaggtc caccgttaca gttggacaca ggccaacttc tcaagattgc aaaacgtggc
      481 ggcgtgaccg cagtggaggc agtgcatgca tggcgcaatg cactgacggg tgccccgctc
      541 aaggagacgg gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
      601 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      661 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      721 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
      781 cgaggccctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
      841 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      901 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      961 cgttgcgctc actgcccgct ttccaccggt cgtctccacc cctgagcagg tagtggctat
     1021 tgcatcccac gacgggggca gacccgcact ggagtcaatt gttgcccagt tatctcgccc
     1081 tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct gcctcggcgg
     1141 gcgtcctgcg ctggatgcag tgaaaaaggg attggggggg tctggatcgt atgctttagg
     1201 accgtatcag atctcagctc cacaattgcc tgcatataac ggacaaactg ttgggacctt
     1261 ttactacgtt aacgatgctg gtggattgga gtccaaagtg ttctcttctg gtggcccaac
     1321 tccatatccc aattatgcga atgcaggcca tgttgaaggt caatcagccc tattcatgag
     1381 agataacgga ataagtgaag gactggtgtt tcacaacaat ccagaaggta cttgtggatt
     1441 ttgcgtaaac atgactgaga cacttctccc agaaaatgcc aagatgacag ttgtacctcc
     1501 tgaaggttct ggtggatcga caaacctttc agacattatc gagaaagaga caggcaaaca
     1561 gctagtgatt caagagtcca ttctcatgct tcccgaagaa gttgaggaag tcattgggaa
     1621 taagccggaa agtgacatac tcgttcatac ggcttacgat gagagcacgg atgagaatgt
     1681 catgttgctt accagtgatg cacctgaata caaaccttgg gctttagtca tccaggacag
     1741 caatggtgag aacaagatca agatgctgtg ataactcgag cgatcctcta gaacccagct
     1801 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1861 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagggcg gccgcgatat
     1921 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1981 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2041 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2101 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2161 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2221 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2281 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2341 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2401 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2461 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gccttgctca
     2521 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2581 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2641 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2701 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2761 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2821 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2881 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2941 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3001 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3061 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3721 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3781 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3841 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3901 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     3961 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     4021 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     4081 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     4141 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     4201 ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat
     4261 tttattt
//
