LOCUS       sequence-379109-        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_NG_G1397-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(16..43)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(135..221)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     285..301
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    318..413
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             416..460
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(770..1180)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1045..1061
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1071..1089
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1137..1157)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1137..1155)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1176..1192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1189..1211)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1200..1216
                     /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(1224..1230,1231..1248,1249..1254))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1269..1290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1854..2105
                     /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(2127..2226)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2257..2275)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2280..2296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2398..2417)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2409..3218
                     /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(2476..2495)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3365..3953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3854..3873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4107..4124
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
       61 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
      121 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
      181 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
      241 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
      301 tcttaagctc gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca
      361 acaaattgat gagcaatgct tttttataat gccaactttg tataataaag ttgataagga
      421 gaccgccgct gccaagttcg agagacagca catggacagc atcgatatcg ccgatctacg
      481 cacgctcggc tacagccagc agcaacagga gaaaatcaaa ccgaaggttc gttcgacagt
      541 ggcgcagcac cacgaggcac tggtcggcca cgggtttaca cacgcgcaca tcgttgcgtt
      601 aagccaacac ccggcagcgt tagggaccgt cgctgtcaag tatcaggaca tgatcgcagc
      661 gttgccagag gcgacacacg aagcgatcgt tggcgtcggc aaacagtggt ccggcgcacg
      721 cgctctggag gccttgctca cggtggcggg agagttgaga ggtccaccgt tacagttgga
      781 cacaggccaa cttctcaaga ttgcaaaacg tggcggcgtg accgcagtgg aggcagtgca
      841 tgcatggcgc aatgcactga cgggtgcccc gctcaaggag acgggcgccg ctacagggcg
      901 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      961 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     1021 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     1081 cactataggg cgaattgggt accgggcccc ccctcgaggt cctccagctt ttgttccctt
     1141 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1201 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1261 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccac
     1321 cggtcgtctc cacccctgag caggtagtgg ctattgcatc caacggaggg ggcagacccg
     1381 cactggagtc aattgttgcc cagttatctc gccctgatcc ggcgttggcc gcgttgacca
     1441 acgaccacct cgtcgccttg gcctgcctcg gcgggcgtcc tgcgctggat gcagtgaaaa
     1501 agggattggg ggggtctgga tcgtatgctt taggaccgta tcagatctca gctccacaat
     1561 tgcctgcata taacggacaa actgttggga ccttttacta cgttaacgat gctggtggat
     1621 tggagtccaa agtgttcatc tctggtggcc caactccata tcccaattat gtgagtgcag
     1681 gccatgttga aggtcaatca gccctattca tgagagataa cggaataagt gaaggactgg
     1741 tgtttcacaa caatccaaaa ggtacttgtg gattttgcgt aaacatgatt gagacacttc
     1801 tcccagaaaa tgccaagatg acagttgtac ctcctgaagg ttctggtgga tcgacaaacc
     1861 tttcagacat tatcgagaaa gagacaggca aacagctagt gattcaagag tccattctca
     1921 tgcttcccga agaagttgag gaagtcattg ggaataagcc ggaaagtgac atactcgttc
     1981 atacggctta cgatgagagc acggatgaga atgtcatgtt gcttaccagt gatgcacctg
     2041 aatacaaacc ttgggcttta gtcatccagg acagcaatgg tgagaacaag atcaagatgc
     2101 tgtgataact cgagcgatcc tctagaaccc agctttcttg tacaaagttg gcattataag
     2161 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2221 tatttgccat ccagctgcag ggcggccgcg atatccccta tagtgagtcg tattacatgg
     2281 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2341 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2401 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2461 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2521 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2581 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2641 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2701 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2761 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2821 cttttaacag cgatcgcgta tttcgccttg ctcaggcgca atcacgaatg aataacggtt
     2881 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2941 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     3001 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     3061 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     3121 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     3181 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     3241 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3301 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3481 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4021 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4081 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4141 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4201 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     4261 aggaaga
//
