LOCUS       sequence-338620-        4267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 1 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_NG_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             463..714
                     /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(736..835)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(866..884)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(889..905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1007..1026)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1018..1827
                     /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(1085..1104)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1974..2562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2463..2482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2716..2733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2892..2919)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3011..3097)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3161..3177
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3194..3289
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3292..3336
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(3646..4056)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3921..3937
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3947..3965
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4013..4033)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4013..4031)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4052..4068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4065..4087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4076..4092
                     /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(4100..4106,4107..4124,4125..4130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4145..4166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
       61 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
      121 gggtctggat cgtatgcttt aggaccgtat cagatctcag ctccacaatt gcctgcatat
      181 aacggacaaa ctgttgggac cttttactac gttaacgatg ctggtggatt ggagtccaaa
      241 gtgttctctt ctggtggccc aactccatat cccaattatg cgaatgcagg ccatgttgaa
      301 ggtcaatcag ccctattcat gagagataac ggaataagtg aaggactggt gtttcacaac
      361 aatccagaag gtacttgtgg attttgcgta aacatgactg agacacttct cccagaaaat
      421 gccaagatga cagttgtacc tcctgaaggt tctggtggat cgacaaacct ttcagacatt
      481 atcgagaaag agacaggcaa acagctagtg attcaagagt ccattctcat gcttcccgaa
      541 gaagttgagg aagtcattgg gaataagccg gaaagtgaca tactcgttca tacggcttac
      601 gatgagagca cggatgagaa tgtcatgttg cttaccagtg atgcacctga atacaaacct
      661 tgggctttag tcatccagga cagcaatggt gagaacaaga tcaagatgct gtgataactc
      721 gagcgatcct ctagaaccca gctttcttgt acaaagttgg cattataaga aagcattgct
      781 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
      841 cagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt
      901 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
      961 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     1021 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     1081 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     1141 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     1201 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     1261 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     1321 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     1381 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     1441 gatcgcgtat ttcgccttgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     1501 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     1561 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     1621 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     1681 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     1741 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     1801 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     1861 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     1921 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2101 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2701 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2761 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2821 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     2881 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     2941 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     3001 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     3061 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     3121 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
     3181 aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa
     3241 attgatgagc aatgcttttt tataatgcca actttgtata ataaagttga taaggagacc
     3301 gccgctgcca agttcgagag acagcacatg gacagcatcg atatcgccga tctacgcacg
     3361 ctcggctaca gccagcagca acaggagaag atcaaaccga aggttcgttc gacagtggcg
     3421 cagcaccacg aggcactggt cggccacggg tttacacacg cgcacatcgt tgcgttaagc
     3481 caacacccgg cagcgttagg gaccgtcgct gtcaagtatc aggacatgat cgcagcgttg
     3541 ccagaggcga cacacgaagc gatcgttggc gtcggcaaac agtggtccgg cgcacgcgct
     3601 ctggaggcct tgctcacggt ggcgggagag ttgagaggtc caccgttaca gttggacaca
     3661 ggccaacttc tcaagattgc aaaacgtggc ggcgtgaccg cagtggaggc agtgcatgca
     3721 tggcgcaatg cactgacggg tgccccgctc aaggagacgg gcgccgctac agggcgcgtc
     3781 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3841 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3901 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     3961 atagggcgaa ttgggtaccg ggccccccct cgaggtcctc cagcttttgt tccctttagt
     4021 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4081 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccaccggt
     4201 cgtctccacc cctgagcagg tagtggctat tgcatccaac ggagggggca gacccgcact
     4261 ggagtca
//
