LOCUS       sequence-338619-        4052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 1 with attL1 and L4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4052)
  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 4052)
  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: E1_pENTR_L1-L4_NN_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             331..582
                     /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(608..703)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(729..747)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(752..768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(870..889)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             881..1690
                     /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(948..967)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1837..2425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2326..2345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2579..2596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2755..2782)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2874..2960)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3024..3040
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3056..3155
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3158..3184
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3491..3901)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3766..3782
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3792..3810
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3858..3878)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3858..3876)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3897..3913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3910..3932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3921..3937
                     /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(3945..3951,3952..3969,3970..3975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3990..4011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 acccctgagc aggtagtggc tattgcatcc aacaacgggg gcagacccgc actggagtca
       61 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
      121 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
      181 ggatctggta gctatgcgtt aggaccctat cagatttcag ctcctcaatt gcctgcctat
      241 aatgggcaaa ctgttggcac cttttactac gtcaatgatg ctggagggtt agaatccaag
      301 gtgttctcaa gtggtggttc tggaggtagt acgaatcttt cggacatcat agagaaggaa
      361 actggaaaac agctcgttat ccaagagagc attctcatgt tgccagaaga agttgaagag
      421 gttataggca acaaaccgga atctgacatt ctggtacata ccgcttatga tgagtcaaca
      481 gatgagaacg tcatgctttt gacatctgat gcaccagaat acaaaccttg ggcacttgtg
      541 attcaggatt ccaatggtga gaacaagatc aagatgctat gataactcga gcgatcctct
      601 agacacccaa cttttctata caaagttggc attataagaa agcattgctt atcaatttgt
      661 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcaggg
      721 cggccgcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
      781 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
      841 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
      901 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
      961 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     1021 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1081 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1141 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     1201 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1261 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgcct
     1321 tgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1381 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     1441 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     1501 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     1561 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     1621 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     1681 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     1741 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     1801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1981 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2221 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2281 aagcggcatg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2581 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2641 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2701 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     2761 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     2821 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     2881 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     2941 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     3001 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat
     3061 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt
     3121 ttttataatg ccaactttgt acaaaaaagc aggcttttac ccatacgatg ttccagatta
     3181 cgctgatatc gccgatctac gcacgctcgg ctacagccag cagcaacagg agaagatcaa
     3241 accgaaggtt cgttcgacag tggcgcagca ccacgaggca ctggtcggcc acgggtttac
     3301 acacgcgcac atcgttgcgt taagccaaca cccggcagcg ttagggaccg tcgctgtcaa
     3361 gtatcaggac atgatcgcag cgttgccaga ggcgacacac gaagcgatcg ttggcgtcgg
     3421 caaacagtgg tccggcgcac gcgctctgga ggccttgctc acggtggcgg gagagttgag
     3481 aggtccaccg ttacagttgg acacaggcca acttctcaag attgcaaaac gtggcggcgt
     3541 gaccgcagtg gaggcagtgc atgcatggcg caatgcactg acgggtgccc cgctcaagga
     3601 gacgggcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     3661 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3721 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3781 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg
     3841 tcctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     3901 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3961 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4021 cgctcactgc ccgctttcca ccggtcgtct cc
//
