LOCUS       sequence-379224-        4244 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 4244)
  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 4244)
  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_NG_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     449..466
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(625..652)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(744..830)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     894..910
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    926..1025
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1028..1054
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1361..1771)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1636..1652
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1662..1680
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1728..1748)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1728..1746)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1767..1783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1780..1802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1791..1807
                     /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(1815..1821,1822..1839,1840..1845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1860..1881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2445..2696
                     /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(2722..2817)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2843..2861)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2866..2882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2984..3003)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2995..3804
                     /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(3062..3081)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3951..4244,1..295)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
       61 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      121 cgaagggaga aaggcggaca ggtatccggt aagcggcatg gtcggaacag gagagcgcac
      181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      361 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      421 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      481 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      541 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
      601 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
      661 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
      721 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      781 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
      841 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
      901 gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg
      961 ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc
     1021 aggcttttac ccatacgatg ttccagatta cgctgatatc gccgatctac gcacgctcgg
     1081 ctacagccag cagcaacagg agaagatcaa accgaaggtt cgttcgacag tggcgcagca
     1141 ccacgaggca ctggtcggcc acgggtttac acacgcgcac atcgttgcgt taagccaaca
     1201 cccggcagcg ttagggaccg tcgctgtcaa gtatcaggac atgatcgcag cgttgccaga
     1261 ggcgacacac gaagcgatcg ttggcgtcgg caaacagtgg tccggcgcac gcgctctgga
     1321 ggccttgctc acggtggcgg gagagttgag aggtccaccg ttacagttgg acacaggcca
     1381 acttctcaag attgcaaaac gtggcggcgt gaccgcagtg gaggcagtgc atgcatggcg
     1441 caatgcactg acgggtgccc cgctcaagga gacgggcgcc gctacagggc gcgtcccatt
     1501 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     1561 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     1621 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     1681 gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct ttagtgaggg
     1741 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1801 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1861 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtcgtct
     1921 ccacccctga gcaggtagtg gctattgcat ccaacggagg gggcagaccc gcactggagt
     1981 caattgttgc ccagttatct cgccctgatc cggcgttggc cgcgttgacc aacgaccacc
     2041 tcgtcgcctt ggcctgcctc ggcgggcgtc ctgcgctgga tgcagtgaaa aagggattgg
     2101 gggggtctgg atcgtatgct ttaggaccgt atcagatctc agctccacaa ttgcctgcat
     2161 ataacggaca aactgttggg accttttact acgttaacga tgctggtgga ttggagtcca
     2221 aagtgttctc ttctggtggc ccaactccat atcccaatta tgcgaatgca ggccatgttg
     2281 aaggtcaatc agccctattc atgagagata acggaataag tgaaggactg gtgtttcaca
     2341 acaatccaga aggtacttgt ggattttgcg taaacatgac tgagacactt ctcccagaaa
     2401 atgccaagat gacagttgta cctcctgaag gttctggtgg atcgacaaac ctttcagaca
     2461 ttatcgagaa agagacaggc aaacagctag tgattcaaga gtccattctc atgcttcccg
     2521 aagaagttga ggaagtcatt gggaataagc cggaaagtga catactcgtt catacggctt
     2581 acgatgagag cacggatgag aatgtcatgt tgcttaccag tgatgcacct gaatacaaac
     2641 cttgggcttt agtcatccag gacagcaatg gtgagaacaa gatcaagatg ctgtgataac
     2701 tcgagcgatc ctctagacac ccaacttttc tatacaaagt tggcattata agaaagcatt
     2761 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2821 atccagctgc agggcggccg cccctatagt gagtcgtatt acatggtcat agctgtttcc
     2881 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     2941 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     3001 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     3061 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     3121 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     3181 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     3241 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     3301 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     3361 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     3421 cgcgtatttc gccttgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     3481 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     3541 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     3601 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     3661 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     3721 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3781 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3841 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     3901 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3961 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4021 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4081 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4141 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4201 gataagtcgt gtcttaccgg gttggactca agacgatagt tacc
//
