LOCUS       sequence-379079-        4075 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 4075)
  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 4075)
  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_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     123..142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     376..393
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(552..579)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(671..757)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     821..837
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    854..949
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             952..996
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1306..1716)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1581..1597
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1607..1625
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1673..1693)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1673..1691)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1712..1728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1725..1747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1736..1752
                     /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(1760..1766,1767..1784,1785..1790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1805..1826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2198..2449
                     /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(2471..2570)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2601..2619)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2624..2640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2742..2761)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2753..3562
                     /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(2820..2839)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3709..4075,1..222)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
       61 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      301 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      361 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      421 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      481 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
      541 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
      601 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
      661 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
      721 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
      781 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
      841 aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa
      901 attgatgagc aatgcttttt tataatgcca actttgtata ataaagttga taaggagacc
      961 gccgctgcca agttcgagag acagcacatg gacagcatcg atatcgccga tctacgcacg
     1021 ctcggctaca gccagcagca acaggagaag atcaaaccga aggttcgttc gacagtggcg
     1081 cagcaccacg aggcactggt cggccacggg tttacacacg cgcacatcgt tgcgttaagc
     1141 caacacccgg cagcgttagg gaccgtcgct gtcaagtatc aggacatgat cgcagcgttg
     1201 ccagaggcga cacacgaagc gatcgttggc gtcggcaaac agtggtccgg cgcacgcgct
     1261 ctggaggcct tgctcacggt ggcgggagag ttgagaggtc caccgttaca gttggacaca
     1321 ggccaacttc tcaagattgc aaaacgtggc ggcgtgaccg cagtggaggc agtgcatgca
     1381 tggcgcaatg cactgacggg tgccccgctc aaggagacgg gcgccgctac agggcgcgtc
     1441 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     1501 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     1561 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     1621 atagggcgaa ttgggtaccg ggccccccct cgaggccctc cagcttttgt tccctttagt
     1681 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccaccggt
     1861 cgtctccacc cctgagcagg tagtggctat tgcatcccac gacgggggca gacccgcact
     1921 ggagtcaatt gttgcccagt tatctcgccc tgatccggcg ttggccgcgt tgaccaacga
     1981 ccacctcgtc gccttggcct gcctcggcgg gcgtcctgcg ctggatgcag tgaaaaaggg
     2041 attgggggga tctggtagct atgcgttagg accctatcag atttcagctc ctcaattgcc
     2101 tgcctataat gggcaaactg ttggcacctt ttactacgtc aatgatgctg gagggttaga
     2161 atccaaggtg ttctcaagtg gtggttctgg aggtagtacg aatctttcgg acatcataga
     2221 gaaggaaact ggaaaacagc tcgttatcca agagagcatt ctcatgttgc cagaagaagt
     2281 tgaagaggtt ataggcaaca aaccggaatc tgacattctg gtacataccg cttatgatga
     2341 gtcaacagat gagaacgtca tgcttttgac atctgatgca ccagaataca aaccttgggc
     2401 acttgtgatt caggattcca atggtgagaa caagatcaag atgctatgat aactcgagcg
     2461 atcctctaga acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     2521 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     2581 gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2641 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2701 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2761 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2821 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2881 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2941 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3001 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3061 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3121 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3181 cgtatttcgc cttgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3241 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3301 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3361 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3421 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3481 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3541 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3601 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3661 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3721 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3781 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3841 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3901 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3961 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4021 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acacc
//
