LOCUS       sequence-379232-        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_HD_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..107
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             110..136
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(443..853)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     718..734
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        744..762
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(810..830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(810..828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(849..865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(862..884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    873..889
                     /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(897..903,904..921,922..927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    942..963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1335..1586
                     /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(1612..1707)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1733..1751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1756..1772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1874..1893)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1885..2694
                     /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(1952..1971)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2841..3429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3330..3349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3583..3600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3759..3786)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3878..3964)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4028..4044
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
       61 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctttt acccatacga
      121 tgttccagat tacgctgata tcgccgatct acgcacgctc ggctacagcc agcagcaaca
      181 ggagaagatc aaaccgaagg ttcgttcgac agtggcgcag caccacgagg cactggtcgg
      241 ccacgggttt acacacgcgc acatcgttgc gttaagccaa cacccggcag cgttagggac
      301 cgtcgctgtc aagtatcagg acatgatcgc agcgttgcca gaggcgacac acgaagcgat
      361 cgttggcgtc ggcaaacagt ggtccggcgc acgcgctctg gaggccttgc tcacggtggc
      421 gggagagttg agaggtccac cgttacagtt ggacacaggc caacttctca agattgcaaa
      481 acgtggcggc gtgaccgcag tggaggcagt gcatgcatgg cgcaatgcac tgacgggtgc
      541 cccgctcaag gagacgggcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      601 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      661 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      721 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
      781 cccccctcga ggccctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
      841 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      901 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      961 ttaattgcgt tgcgctcact gcccgctttc caccggtcgt ctccacccct gagcaggtag
     1021 tggctattgc atcccacgac gggggcagac ccgcactgga gtcaattgtt gcccagttat
     1081 ctcgccctga tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc
     1141 tcggcgggcg tcctgcgctg gatgcagtga aaaagggatt ggggggatct ggtagctatg
     1201 cgttaggacc ctatcagatt tcagctcctc aattgcctgc ctataatggg caaactgttg
     1261 gcacctttta ctacgtcaat gatgctggag ggttagaatc caaggtgttc tcaagtggtg
     1321 gttctggagg tagtacgaat ctttcggaca tcatagagaa ggaaactgga aaacagctcg
     1381 ttatccaaga gagcattctc atgttgccag aagaagttga agaggttata ggcaacaaac
     1441 cggaatctga cattctggta cataccgctt atgatgagtc aacagatgag aacgtcatgc
     1501 ttttgacatc tgatgcacca gaatacaaac cttgggcact tgtgattcag gattccaatg
     1561 gtgagaacaa gatcaagatg ctatgataac tcgagcgatc ctctagacac ccaacttttc
     1621 tatacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1681 ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg cccctatagt
     1741 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     1801 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     1861 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     1921 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1981 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     2041 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     2101 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     2161 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     2221 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     2281 cctgtttgta attgtccttt taacagcgat cgcgtatttc gccttgctca ggcgcaatca
     2341 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     2401 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     2461 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     2521 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     2581 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     2641 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     2701 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     2761 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     2821 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2881 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2941 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3001 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3061 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg catggtcgga
     3301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3481 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3541 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3601 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3661 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3721 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     3781 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     3841 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     3901 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     3961 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     4021 cgacgttgta aaacgacggc cagtcttaag ct
//
