LOCUS       sequence-393337-        4736 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 4736)
  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 4736)
  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_HD_G1333-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(388..415)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(507..593)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     657..673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    690..785
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             788..832
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1142..1552)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1417..1433
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1443..1461
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1509..1529)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1509..1527)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1548..1564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1561..1583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1572..1588
                     /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(1596..1602,1603..1620,1621..1626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1641..1662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2034..2285
                     /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(2307..2406)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2437..2455)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2460..2476)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2578..2597)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2589..3398
                     /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(2656..2675)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3545..4133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4034..4053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  4438..4662
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      241 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      301 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      361 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      421 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      481 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      541 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      601 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      661 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      721 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa
      781 agttgataag gagaccgccg ctgccaagtt cgagagacag cacatggaca gcatcgatat
      841 cgccgatcta cgcacgctcg gctacagcca gcagcaacag gagaagatca aaccgaaggt
      901 tcgttcgaca gtggcgcagc accacgaggc actggtcggc cacgggttta cacacgcgca
      961 catcgttgcg ttaagccaac acccggcagc gttagggacc gtcgctgtca agtatcagga
     1021 catgatcgca gcgttgccag aggcgacaca cgaagcgatc gttggcgtcg gcaaacagtg
     1081 gtccggcgca cgcgctctgg aggccttgct cacggtggcg ggagagttga gaggtccacc
     1141 gttacagttg gacacaggcc aacttctcaa gattgcaaaa cgtggcggcg tgaccgcagt
     1201 ggaggcagtg catgcatggc gcaatgcact gacgggtgcc ccgctcaagg agacgggcgc
     1261 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1321 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1381 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     1441 cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gccctccagc
     1501 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     1561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1681 cccgctttcc accggtcgtc tccacccctg agcaggtagt ggctattgca tcccacgacg
     1741 ggggcagacc cgcactggag tcaattgttg cccagttatc tcgccctgat ccggcgttgg
     1801 ccgcgttgac caacgaccac ctcgtcgcct tggcctgcct cggcgggcgt cctgcgctgg
     1861 atgcagtgaa aaagggattg gggggatctg gtagctatgc gttaggaccc tatcagattt
     1921 cagctcctca attgcctgcc tataatgggc aaactgttgg caccttttac tacgtcaatg
     1981 atgctggagg gttagaatcc aaggtgttca taagtggtgg ttctggaggt agtacgaatc
     2041 tttcggacat catagagaag gaaactggaa aacagctcgt tatccaagag agcattctca
     2101 tgttgccaga agaagttgaa gaggttatag gcaacaaacc ggaatctgac attctggtac
     2161 ataccgctta tgatgagtca acagatgaga acgtcatgct tttgacatct gatgcaccag
     2221 aatacaaacc ttgggcactt gtgattcagg attccaatgg tgagaacaag atcaagatgc
     2281 tatgataact cgagcgatcc tctagaaccc agctttcttg tacaaagttg gcattataag
     2341 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2401 tatttgccat ccagctgcag ggcggccgcg atatccccta tagtgagtcg tattacatgg
     2461 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2521 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2581 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2641 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2701 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2761 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2821 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2881 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2941 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     3001 cttttaacag cgatcgcgta tttcgccttg ctcaggcgca atcacgaatg aataacggtt
     3061 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     3121 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     3181 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     3241 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     3301 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     3361 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     3421 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3481 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3541 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4141 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4201 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4261 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4321 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gaccagccgc
     4381 gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt aagcgggtgt
     4441 gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca ataaagtctt
     4501 aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa aaagcatact
     4561 ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt gcccgtcgta
     4621 ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt gtgacaattt
     4681 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtgg
//
