LOCUS       sequence-393338-        4075 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 4075)
  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 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: "10.1111/tpj.16311";
            journalName: "Plant J"; date: "2023-08"; volume: "115"; issue: "4"; 
            pages: "1151-1162"
COMMENT     Sequence Label: E3_pENTR_L3-L2_NN_G1333-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(12..98)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     162..178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    195..290
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             293..337
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(647..1057)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     922..938
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        948..966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1014..1034)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1014..1032)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1053..1069)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1066..1088)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1077..1093
                     /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(1101..1107,1108..1125,1126..1131))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1146..1167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1539..1790
                     /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(1812..1911)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1942..1960)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1965..1981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2083..2102)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2094..2903
                     /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(2161..2180)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3050..3638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3539..3558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3792..3809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3968..3995)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
       61 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      121 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct
      181 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
      241 aattgatgag caatgctttt ttataatgcc aactttgtat aataaagttg ataaggagac
      301 cgccgctgcc aagttcgaga gacagcacat ggacagcatc gatatcgccg atctacgcac
      361 gctcggctac agccagcagc aacaggagaa gatcaaaccg aaggttcgtt cgacagtggc
      421 gcagcaccac gaggcactgg tcggccacgg gtttacacac gcgcacatcg ttgcgttaag
      481 ccaacacccg gcagcgttag ggaccgtcgc tgtcaagtat caggacatga tcgcagcgtt
      541 gccagaggcg acacacgaag cgatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc
      601 tctggaggcc ttgctcacgg tggcgggaga gttgagaggt ccaccgttac agttggacac
      661 aggccaactt ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc
      721 atggcgcaat gcactgacgg gtgccccgct caaggagacg ggcgccgcta cagggcgcgt
      781 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      841 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      901 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
      961 tatagggcga attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag
     1021 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1081 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1141 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg
     1201 tcgtctccac ccctgagcag gtagtggcta ttgcatccaa caacgggggc agacccgcac
     1261 tggagtcaat tgttgcccag ttatctcgcc ctgatccggc gttggccgcg ttgaccaacg
     1321 accacctcgt cgccttggcc tgcctcggcg ggcgtcctgc gctggatgca gtgaaaaagg
     1381 gattgggggg atctggtagc tatgcgttag gaccctatca gatttcagct cctcaattgc
     1441 ctgcctataa tgggcaaact gttggcacct tttactacgt caatgatgct ggagggttag
     1501 aatccaaggt gttcataagt ggtggttctg gaggtagtac gaatctttcg gacatcatag
     1561 agaaggaaac tggaaaacag ctcgttatcc aagagagcat tctcatgttg ccagaagaag
     1621 ttgaagaggt tataggcaac aaaccggaat ctgacattct ggtacatacc gcttatgatg
     1681 agtcaacaga tgagaacgtc atgcttttga catctgatgc accagaatac aaaccttggg
     1741 cacttgtgat tcaggattcc aatggtgaga acaagatcaa gatgctatga taactcgagc
     1801 gatcctctag aacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     1861 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     1921 tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     1981 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     2041 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2101 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     2161 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     2221 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2281 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2341 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     2401 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     2461 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2521 gcgtatttcg ccttgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2581 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     2641 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2701 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2761 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2821 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2881 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     2941 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     3001 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3061 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3121 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3181 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3661 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3721 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3781 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3901 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     3961 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     4021 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatcc
//
