LOCUS       sequence-379210-        4267 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 4267)
  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 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: 
            "10.1038/s41477-021-00954-6"; journalName: "Nat Plants"; date: 
            "2021-07"; volume: "7"; issue: "7"; pages: "906-913"
COMMENT     Sequence Label: E3_pENTR_L3-L2_NI_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(147..174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(266..352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     416..432
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    449..544
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             547..591
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(901..1311)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1176..1192
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1202..1220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1268..1288)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1268..1286)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1307..1323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1320..1342)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1331..1347
                     /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(1355..1361,1362..1379,1380..1385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1400..1421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1985..2236
                     /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(2258..2357)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2388..2406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2411..2427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2529..2548)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2540..3349
                     /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(2607..2626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3985..4004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4238..4255
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      121 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
      241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
      301 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
      361 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
      421 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
      481 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtataataaa
      541 gttgataagg agaccgccgc tgccaagttc gagagacagc acatggacag catcgatatc
      601 gccgatctac gcacgctcgg ctacagccag cagcaacagg agaagatcaa accgaaggtt
      661 cgttcgacag tggcgcagca ccacgaggca ctggtcggcc acgggtttac acacgcgcac
      721 atcgttgcgt taagccaaca cccggcagcg ttagggaccg tcgctgtcaa gtatcaggac
      781 atgatcgcag cgttgccaga ggcgacacac gaagcgatcg ttggcgtcgg caaacagtgg
      841 tccggcgcac gcgctctgga ggccttgctc acggtggcgg gagagttgag aggtccaccg
      901 ttacagttgg acacaggcca acttctcaag attgcaaaac gtggcggcgt gaccgcagtg
      961 gaggcagtgc atgcatggcg caatgcactg acgggtgccc cgctcaagga gacgggcgcc
     1021 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     1081 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     1141 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
     1201 gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg tcctccagct
     1261 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     1321 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1381 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1441 ccgctttcca ccggtcgtct ccacccctga gcaggtagtg gctattgcat ccaacatagg
     1501 gggcagaccc gcactggagt caattgttgc ccagttatct cgccctgatc cggcgttggc
     1561 cgcgttgacc aacgaccacc tcgtcgcctt ggcctgcctc ggcgggcgtc ctgcgctgga
     1621 tgcagtgaaa aagggattgg gggggtctgg atcgtatgct ttaggaccgt atcagatctc
     1681 agctccacaa ttgcctgcat ataacggaca aactgttggg accttttact acgttaacga
     1741 tgctggtgga ttggagtcca aagtgttctc ttctggtggc ccaactccat atcccaatta
     1801 tgcgaatgca ggccatgttg aaggtcaatc agccctattc atgagagata acggaataag
     1861 tgaaggactg gtgtttcaca acaatccaga aggtacttgt ggattttgcg taaacatgac
     1921 tgagacactt ctcccagaaa atgccaagat gacagttgta cctcctgaag gttctggtgg
     1981 atcgacaaac ctttcagaca ttatcgagaa agagacaggc aaacagctag tgattcaaga
     2041 gtccattctc atgcttcccg aagaagttga ggaagtcatt gggaataagc cggaaagtga
     2101 catactcgtt catacggctt acgatgagag cacggatgag aatgtcatgt tgcttaccag
     2161 tgatgcacct gaatacaaac cttgggcttt agtcatccag gacagcaatg gtgagaacaa
     2221 gatcaagatg ctgtgataac tcgagcgatc ctctagaacc cagctttctt gtacaaagtt
     2281 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2341 aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct atagtgagtc
     2401 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2461 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2521 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2581 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2641 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2701 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2761 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2821 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2881 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2941 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctt gctcaggcgc aatcacgaat
     3001 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3061 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3121 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3181 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccattctat ggaactgcct
     3241 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3301 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3361 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3421 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4201 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4261 ggaagag
//
