LOCUS       sequence-379067-        4225 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 4225)
  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 4225)
  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_HD_G1333-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(64..150)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     214..230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    247..342
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             345..389
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(699..1109)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     974..990
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1000..1018
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1066..1086)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1066..1084)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1105..1121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1118..1140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1129..1145
                     /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(1153..1159,1160..1177,1178..1183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1198..1219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1741..1992
                     /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(2014..2113)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2144..2162)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2167..2183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2285..2304)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2296..3105
                     /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(2363..2382)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3252..3840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3741..3760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3994..4011
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4170..4197)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
       61 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      121 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      181 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      241 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      301 agcaatgctt ttttataatg ccaactttgt ataataaagt tgataaggag accgccgctg
      361 ccaagttcga gagacagcac atggacagca tcgatatcgc cgatctacgc acgctcggct
      421 acagccagca gcaacaggag aagatcaaac cgaaggttcg ttcgacagtg gcgcagcacc
      481 acgaggcact ggtcggccac gggtttacac acgcgcacat cgttgcgtta agccaacacc
      541 cggcagcgtt agggaccgtc gctgtcaagt atcaggacat gatcgcagcg ttgccagagg
      601 cgacacacga agcgatcgtt ggcgtcggca aacagtggtc cggcgcacgc gctctggagg
      661 ccttgctcac ggtggcggga gagttgagag gtccaccgtt acagttggac acaggccaac
      721 ttctcaagat tgcaaaacgt ggcggcgtga ccgcagtgga ggcagtgcat gcatggcgca
      781 atgcactgac gggtgccccg ctcaaggaga cgggcgccgc tacagggcgc gtcccattcg
      841 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      901 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      961 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     1021 gaattgggta ccgggccccc cctcgaggcc ctccagcttt tgttcccttt agtgagggtt
     1081 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1141 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1201 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccacc ggtcgtctcc
     1261 acccctgagc aggtagtggc tattgcatcc cacgacgggg gcagacccgc actggagtca
     1321 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
     1381 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
     1441 ggtagtccaa ctccgtatcc gaattacgcc aatgcaggac atgttgaagg tcaatctgca
     1501 ttgttcatga gggataacgg catttctgaa gggttggtgt tccacaacaa ccctgaagga
     1561 acatgtggat tttgcgtcaa catgacagaa acccttctcc cagaaaacgc taagatgaca
     1621 gtagttccac ctgaaggtgc tattcctgtc aaaagaggtg ctactggtga aaccaaggtg
     1681 tttactggga attccaattc acccaaaagc ccaacgaaag gtgggtgtag tggaggatct
     1741 acaaatctct ctgacatcat tgagaaagag actggaaagc aactagtcat tcaggagtca
     1801 atcctgatgt taccagagga ggttgaggaa gtgataggca ataagcccga aagcgatata
     1861 cttgttcata ctgcctatga cgaatcgacg gatgagaacg taatgcttct aacctcagat
     1921 gctcctgagt acaaaccttg ggcgttagtt atccaggatt ccaatggaga gaacaagatc
     1981 aagatgttgt gataactcga gcgatcctct agaacccagc tttcttgtac aaagttggca
     2041 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2101 aaatcattat ttgccatcca gctgcagggc ggccgcgata tcccctatag tgagtcgtat
     2161 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2221 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2281 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2341 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2401 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2461 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2521 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2581 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2641 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     2701 aattgtcctt ttaacagcga tcgcgtattt cgccttgctc aggcgcaatc acgaatgaat
     2761 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2821 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2881 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2941 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3001 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3061 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3121 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3181 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3241 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3301 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3361 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3421 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3481 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3541 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3601 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3661 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3721 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3781 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3841 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3901 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3961 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4021 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4081 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     4141 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     4201 tagtttgatg cctggcagtt tatgg
//
