LOCUS       sequence-379107-        4267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD_v2 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 4267)
  AUTHORS   Nakazato I, Okuno M, Itoh T, Tsutsumi N, Arimura SI
  TITLE     Characterization and development of a plastid genome base editor, 
            ptpTALECD.
  JOURNAL   Plant J. 2023 Aug;115(4):1151-1162. doi: 10.1111/tpj.16311. Epub 
            2023 Jun 2.
  PUBMED    37265080
REFERENCE   2  (bases 1 to 4267)
  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: "10.1111/tpj.16311";
            journalName: "Plant J"; date: "2023-08"; volume: "115"; issue: "4"; 
            pages: "1151-1162"
COMMENT     Sequence Label: E3_pENTR_L3-L2_NN_G1397-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..28
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     262..279
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(438..465)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(557..643)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     707..723
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    740..835
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             838..882
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1192..1602)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1467..1483
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1493..1511
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1559..1579)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1559..1577)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1598..1614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1611..1633)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1622..1638
                     /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(1646..1652,1653..1670,1671..1676))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1691..1712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2276..2527
                     /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(2549..2648)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2679..2697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2702..2718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2820..2839)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2831..3640
                     /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(2898..2917)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3787..4267,1..108)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
       61 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      121 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      181 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      241 cacagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
      301 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
      361 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
      421 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
      481 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
      541 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
      601 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
      661 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
      721 agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg
      781 caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa agttgataag
      841 gagaccgccg ctgccaagtt cgagagacag cacatggaca gcatcgatat cgccgatcta
      901 cgcacgctcg gctacagcca gcagcaacag gagaagatca aaccgaaggt tcgttcgaca
      961 gtggcgcagc accacgaggc actggtcggc cacgggttta cacacgcgca catcgttgcg
     1021 ttaagccaac acccggcagc gttagggacc gtcgctgtca agtatcagga catgatcgca
     1081 gcgttgccag aggcgacaca cgaagcgatc gttggcgtcg gcaaacagtg gtccggcgca
     1141 cgcgctctgg aggccttgct cacggtggcg ggagagttga gaggtccacc gttacagttg
     1201 gacacaggcc aacttctcaa gattgcaaaa cgtggcggcg tgaccgcagt ggaggcagtg
     1261 catgcatggc gcaatgcact gacgggtgcc ccgctcaagg agacgggcgc cgctacaggg
     1321 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1381 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1441 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     1501 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc ttttgttccc
     1561 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1621 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1681 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1741 accggtcgtc tccacccctg agcaggtagt ggctattgca tccaacaacg ggggcagacc
     1801 cgcactggag tcaattgttg cccagttatc tcgccctgat ccggcgttgg ccgcgttgac
     1861 caacgaccac ctcgtcgcct tggcctgcct cggcgggcgt cctgcgctgg atgcagtgaa
     1921 aaagggattg ggggggtctg gatcgtatgc tttaggaccg tatcagatct cagctccaca
     1981 attgcctgca tataacggac aaactgttgg gaccttttac tacgttaacg atgctggtgg
     2041 attggagtcc aaagtgttca tctctggtgg cccaactcca tatcccaatt atgtgagtgc
     2101 aggccatgtt gaaggtcaat cagccctatt catgagagat aacggaataa gtgaaggact
     2161 ggtgtttcac aacaatccaa aaggtacttg tggattttgc gtaaacatga ttgagacact
     2221 tctcccagaa aatgccaaga tgacagttgt acctcctgaa ggttctggtg gatcgacaaa
     2281 cctttcagac attatcgaga aagagacagg caaacagcta gtgattcaag agtccattct
     2341 catgcttccc gaagaagttg aggaagtcat tgggaataag ccggaaagtg acatactcgt
     2401 tcatacggct tacgatgaga gcacggatga gaatgtcatg ttgcttacca gtgatgcacc
     2461 tgaatacaaa ccttgggctt tagtcatcca ggacagcaat ggtgagaaca agatcaagat
     2521 gctgtgataa ctcgagcgat cctctagaac ccagctttct tgtacaaagt tggcattata
     2581 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     2641 attatttgcc atccagctgc agggcggccg cgatatcccc tatagtgagt cgtattacat
     2701 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2761 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2821 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2881 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2941 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3001 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3061 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3121 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3181 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3241 tccttttaac agcgatcgcg tatttcgcct tgctcaggcg caatcacgaa tgaataacgg
     3301 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3361 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3421 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3481 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3541 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3601 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3661 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3721 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggag
//
