LOCUS       sequence-379219-        4033 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 4033)
  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 4033)
  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_G1397-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(563..590)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(682..768)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     832..848
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    865..960
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             963..1007
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1317..1727)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1592..1608
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1618..1636
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1684..1704)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1684..1702)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1723..1739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1736..1758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1747..1763
                     /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(1771..1777,1778..1795,1796..1801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1816..1837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2167..2418
                     /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(2440..2539)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2570..2588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2593..2609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2711..2730)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2722..3531
                     /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(2789..2808)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3678..4033,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
      541 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
      601 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
      661 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      721 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
      781 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
      841 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
      901 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtat aataaagttg
      961 ataaggagac cgccgctgcc aagttcgaga gacagcacat ggacagcatc gatatcgccg
     1021 atctacgcac gctcggctac agccagcagc aacaggagaa gatcaaaccg aaggttcgtt
     1081 cgacagtggc gcagcaccac gaggcactgg tcggccacgg gtttacacac gcgcacatcg
     1141 ttgcgttaag ccaacacccg gcagcgttag ggaccgtcgc tgtcaagtat caggacatga
     1201 tcgcagcgtt gccagaggcg acacacgaag cgatcgttgg cgtcggcaaa cagtggtccg
     1261 gcgcacgcgc tctggaggcc ttgctcacgg tggcgggaga gttgagaggt ccaccgttac
     1321 agttggacac aggccaactt ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg
     1381 cagtgcatgc atggcgcaat gcactgacgg gtgccccgct caaggagacg ggcgccgcta
     1441 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1501 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     1561 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
     1621 tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcct ccagcttttg
     1681 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     1741 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1801 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1861 tttccaccgg tcgtctccac ccctgagcag gtagtggcta ttgcatccaa catagggggc
     1921 agacccgcac tggagtcaat tgttgcccag ttatctcgcc ctgatccggc gttggccgcg
     1981 ttgaccaacg accacctcgt cgccttggcc tgcctcggcg ggcgtcctgc gctggatgca
     2041 gtgaaaaagg gattgggggg ttctgcgatt ccagttaaga gaggagctac aggagaaacg
     2101 aaagtcttta ctgggaattc caattctccc aaatcaccga ctaaaggcgg atgtagtggt
     2161 ggtagtacca atctttccga cattatcgag aaggaaacag gtaaacaact cgtaatccaa
     2221 gaaagcatac tgatgcttcc tgaagaggtt gaagaggtca tagggaacaa acctgaaagc
     2281 gacattttgg ttcatactgc ctatgatgag tctacagatg agaacgtgat gttgctaacc
     2341 tcagatgcac ctgaatacaa gccatgggct ttagtgattc aggattcgaa tggagagaac
     2401 aagatcaaga tgctctgata actcgagcga tcctctagaa cccagctttc ttgtacaaag
     2461 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     2521 aaaataaaat cattatttgc catccagctg cagggcggcc gcgatatccc ctatagtgag
     2581 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     2641 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     2701 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     2761 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2821 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2881 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2941 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     3001 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     3061 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3121 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgcc ttgctcaggc gcaatcacga
     3181 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3241 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3301 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3361 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3421 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3481 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     3541 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     3601 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     3661 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3721 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3781 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3841 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3901 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3961 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4021 agcttggagc gaa
//
