LOCUS       sequence-338633-        4202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 1 with attL1 and L4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4202)
  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 4202)
  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: E1_pENTR_L1-L4_NN_G1333-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             481..732
                     /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(758..853)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(879..897)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(902..918)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1020..1039)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1031..1840
                     /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(1098..1117)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1987..2575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2476..2495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2729..2746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2905..2932)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3024..3110)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3174..3190
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3206..3305
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3308..3334
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3641..4051)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3916..3932
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3942..3960
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4008..4028)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4008..4026)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4047..4063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4060..4082)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4071..4087
                     /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(4095..4101,4102..4119,4120..4125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4140..4161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 acccctgagc aggtagtggc tattgcatcc aacaacgggg gcagacccgc actggagtca
       61 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
      121 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
      181 ggtagtccaa ctccgtatcc gaattacgcc aatgcaggac atgttgaagg tcaatctgca
      241 ttgttcatga gggataacgg catttctgaa gggttggtgt tccacaacaa ccctgaagga
      301 acatgtggat tttgcgtcaa catgacagaa acccttctcc cagaaaacgc taagatgaca
      361 gtagttccac ctgaaggtgc tattcctgtc aaaagaggtg ctactggtga aaccaaggtg
      421 tttactggga attccaattc acccaaaagc ccaacgaaag gtgggtgtag tggaggatct
      481 acaaatctct ctgacatcat tgagaaagag actggaaagc aactagtcat tcaggagtca
      541 atcctgatgt taccagagga ggttgaggaa gtgataggca ataagcccga aagcgatata
      601 cttgttcata ctgcctatga cgaatcgacg gatgagaacg taatgcttct aacctcagat
      661 gctcctgagt acaaaccttg ggcgttagtt atccaggatt ccaatggaga gaacaagatc
      721 aagatgttgt gataactcga gcgatcctct agacacccaa cttttctata caaagttggc
      781 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
      841 aaaatcatta tttgccatcc agctgcaggg cggccgcccc tatagtgagt cgtattacat
      901 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
      961 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1021 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1081 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1141 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1201 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1261 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1321 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1381 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1441 tccttttaac agcgatcgcg tatttcgcct tgctcaggcg caatcacgaa tgaataacgg
     1501 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     1561 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     1621 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     1681 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     1741 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     1801 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     1861 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     1921 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2101 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2401 cgaagggaga aaggcggaca ggtatccggt aagcggcatg gtcggaacag gagagcgcac
     2461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2641 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2701 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2761 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2821 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
     2881 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
     2941 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     3001 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     3061 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
     3121 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
     3181 gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg
     3241 ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc
     3301 aggcttttac ccatacgatg ttccagatta cgctgatatc gccgatctac gcacgctcgg
     3361 ctacagccag cagcaacagg agaagatcaa accgaaggtt cgttcgacag tggcgcagca
     3421 ccacgaggca ctggtcggcc acgggtttac acacgcgcac atcgttgcgt taagccaaca
     3481 cccggcagcg ttagggaccg tcgctgtcaa gtatcaggac atgatcgcag cgttgccaga
     3541 ggcgacacac gaagcgatcg ttggcgtcgg caaacagtgg tccggcgcac gcgctctgga
     3601 ggccttgctc acggtggcgg gagagttgag aggtccaccg ttacagttgg acacaggcca
     3661 acttctcaag attgcaaaac gtggcggcgt gaccgcagtg gaggcagtgc atgcatggcg
     3721 caatgcactg acgggtgccc cgctcaagga gacgggcgcc gctacagggc gcgtcccatt
     3781 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     3841 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     3901 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     3961 gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct ttagtgaggg
     4021 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4081 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4141 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtcgtct
     4201 cc
//
