LOCUS       sequence-338634-        4033 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 4033)
  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 4033)
  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-C
FEATURES             Location/Qualifiers
     source          1..4033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             229..480
                     /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(502..601)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(632..650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(655..671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(773..792)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             784..1593
                     /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(851..870)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1740..2328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2229..2248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2482..2499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2658..2685)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2777..2863)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2927..2943
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2960..3055
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3058..3102
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(3412..3822)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3687..3703
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3713..3731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3779..3799)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3779..3797)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3818..3834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3831..3853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3842..3858
                     /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(3866..3872,3873..3890,3891..3896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3911..3932
                     /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 ggttctgcga ttccagttaa gagaggagct acaggagaaa cgaaagtctt tactgggaat
      181 tccaattctc ccaaatcacc gactaaaggc ggatgtagtg gtggtagtac caatctttcc
      241 gacattatcg agaaggaaac aggtaaacaa ctcgtaatcc aagaaagcat actgatgctt
      301 cctgaagagg ttgaagaggt catagggaac aaacctgaaa gcgacatttt ggttcatact
      361 gcctatgatg agtctacaga tgagaacgtg atgttgctaa cctcagatgc acctgaatac
      421 aagccatggg ctttagtgat tcaggattcg aatggagaga acaagatcaa gatgctctga
      481 taactcgagc gatcctctag aacccagctt tcttgtacaa agttggcatt ataagaaagc
      541 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
      601 gccatccagc tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata
      661 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
      721 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
      781 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
      841 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
      901 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
      961 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1021 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1081 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1141 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1201 aacagcgatc gcgtatttcg ccttgctcag gcgcaatcac gaatgaataa cggtttggtt
     1261 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1321 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1381 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1441 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     1501 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     1561 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     1621 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     1681 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1741 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1801 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1861 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1921 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1981 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2041 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2101 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2161 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2221 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2281 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2341 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2401 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2461 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2521 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2581 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     2641 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     2701 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     2761 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     2821 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     2881 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     2941 agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg
     3001 caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa agttgataag
     3061 gagaccgccg ctgccaagtt cgagagacag cacatggaca gcatcgatat cgccgatcta
     3121 cgcacgctcg gctacagcca gcagcaacag gagaagatca aaccgaaggt tcgttcgaca
     3181 gtggcgcagc accacgaggc actggtcggc cacgggttta cacacgcgca catcgttgcg
     3241 ttaagccaac acccggcagc gttagggacc gtcgctgtca agtatcagga catgatcgca
     3301 gcgttgccag aggcgacaca cgaagcgatc gttggcgtcg gcaaacagtg gtccggcgca
     3361 cgcgctctgg aggccttgct cacggtggcg ggagagttga gaggtccacc gttacagttg
     3421 gacacaggcc aacttctcaa gattgcaaaa cgtggcggcg tgaccgcagt ggaggcagtg
     3481 catgcatggc gcaatgcact gacgggtgcc ccgctcaagg agacgggcgc cgctacaggg
     3541 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     3601 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     3661 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     3721 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc ttttgttccc
     3781 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3841 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3901 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3961 accggtcgtc tccacccctg agcaggtagt ggctattgca tccaacggag ggggcagacc
     4021 cgcactggag tca
//
