LOCUS       sequence-379209-        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_NN_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(140..167)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(259..345)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     409..425
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    442..537
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             540..584
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(894..1304)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1169..1185
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1195..1213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1261..1281)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1261..1279)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1300..1316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1313..1335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1324..1340
                     /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(1348..1354,1355..1372,1373..1378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1393..1414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1978..2229
                     /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(2251..2350)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2404..2420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2522..2541)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2533..3342
                     /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(2600..2619)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3489..4077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3978..3997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4231..4248
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
       61 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
      121 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
      181 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
      241 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
      301 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
      361 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
      421 ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt
      481 tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtataat aaagttgata
      541 aggagaccgc cgctgccaag ttcgagagac agcacatgga cagcatcgat atcgccgatc
      601 tacgcacgct cggctacagc cagcagcaac aggagaagat caaaccgaag gttcgttcga
      661 cagtggcgca gcaccacgag gcactggtcg gccacgggtt tacacacgcg cacatcgttg
      721 cgttaagcca acacccggca gcgttaggga ccgtcgctgt caagtatcag gacatgatcg
      781 cagcgttgcc agaggcgaca cacgaagcga tcgttggcgt cggcaaacag tggtccggcg
      841 cacgcgctct ggaggccttg ctcacggtgg cgggagagtt gagaggtcca ccgttacagt
      901 tggacacagg ccaacttctc aagattgcaa aacgtggcgg cgtgaccgca gtggaggcag
      961 tgcatgcatg gcgcaatgca ctgacgggtg ccccgctcaa ggagacgggc gccgctacag
     1021 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     1081 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     1141 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     1201 gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc
     1261 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1441 ccaccggtcg tctccacccc tgagcaggta gtggctattg catccaacaa cgggggcaga
     1501 cccgcactgg agtcaattgt tgcccagtta tctcgccctg atccggcgtt ggccgcgttg
     1561 accaacgacc acctcgtcgc cttggcctgc ctcggcgggc gtcctgcgct ggatgcagtg
     1621 aaaaagggat tgggggggtc tggatcgtat gctttaggac cgtatcagat ctcagctcca
     1681 caattgcctg catataacgg acaaactgtt gggacctttt actacgttaa cgatgctggt
     1741 ggattggagt ccaaagtgtt ctcttctggt ggcccaactc catatcccaa ttatgcgaat
     1801 gcaggccatg ttgaaggtca atcagcccta ttcatgagag ataacggaat aagtgaagga
     1861 ctggtgtttc acaacaatcc agaaggtact tgtggatttt gcgtaaacat gactgagaca
     1921 cttctcccag aaaatgccaa gatgacagtt gtacctcctg aaggttctgg tggatcgaca
     1981 aacctttcag acattatcga gaaagagaca ggcaaacagc tagtgattca agagtccatt
     2041 ctcatgcttc ccgaagaagt tgaggaagtc attgggaata agccggaaag tgacatactc
     2101 gttcatacgg cttacgatga gagcacggat gagaatgtca tgttgcttac cagtgatgca
     2161 cctgaataca aaccttgggc tttagtcatc caggacagca atggtgagaa caagatcaag
     2221 atgctgtgat aactcgagcg atcctctaga acccagcttt cttgtacaaa gttggcatta
     2281 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2341 tcattatttg ccatccagct gcagggcggc cgcgatatcc cctatagtga gtcgtattac
     2401 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2461 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2521 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2581 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2641 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2701 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2761 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2821 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2881 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2941 tgtcctttta acagcgatcg cgtatttcgc cttgctcagg cgcaatcacg aatgaataac
     3001 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3061 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3121 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3181 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3241 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3301 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3361 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3421 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3601 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3841 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4141 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4201 accgctcgcc acagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4261 cgcccaa
//
