LOCUS       sequence-338460-        4075 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 4075)
  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 4075)
  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_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      10..37
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(196..213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(367..955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(447..466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1102..1911)
                     /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     1825..1844
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1903..1922
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2024..2040
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2045..2063
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2094..2193
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2215..2466)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    2838..2859
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2874..2879,2880..2897,2898..2904)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2912..2928
                     /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)."
     primer_bind     2917..2939
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2936..2952
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2948..3358
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     2971..2991
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2973..2991
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3039..3057)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3067..3083)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3668..3712)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    complement(3715..3810)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3827..3843)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3907..3993
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaactaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttcctg
       61 gctagcggta cgcgtattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      121 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      181 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      241 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      301 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      361 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      421 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      481 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      541 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      601 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      661 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      721 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      781 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      841 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      901 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      961 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgtaac tcacgttaag
     1021 ggattttggt catgagcttg cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta
     1081 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1141 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1201 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1261 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1321 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1381 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1441 accgttattc attcgtgatt gcgcctgagc aaggcgaaat acgcgatcgc tgttaaaagg
     1501 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1561 attttcacct gaatcaggat attcttctaa tacctggaat gctgtttttc cggggatcgc
     1621 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1681 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1741 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat
     1801 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1861 catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca taacacccct
     1921 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     1981 tgcaatgtaa catcagagat tttgagacac gggccagagc tgccaggaaa cagctatgac
     2041 catgtaatac gactcactat aggggatatc gcggccgccc tgcagctgga tggcaaataa
     2101 tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgataag caatgctttc
     2161 ttataatgcc aactttgtac aagaaagctg ggttctagag gatcgctcga gttatcatag
     2221 catcttgatc ttgttctcac cattggaatc ctgaatcaca agtgcccaag gtttgtattc
     2281 tggtgcatca gatgtcaaaa gcatgacgtt ctcatctgtt gactcatcat aagcggtatg
     2341 taccagaatg tcagattccg gtttgttgcc tataacctct tcaacttctt ctggcaacat
     2401 gagaatgctc tcttggataa cgagctgttt tccagtttcc ttctctatga tgtccgaaag
     2461 attcgtacta cctccagaac caccacttga gaacaccttg gattctaacc ctccagcatc
     2521 attgacgtag taaaaggtgc caacagtttg cccattatag gcaggcaatt gaggagctga
     2581 aatctgatag ggtcctaacg catagctacc agatcccccc aatccctttt tcactgcatc
     2641 cagcgcagga cgcccgccga ggcaggccaa ggcgacgagg tggtcgttgg tcaacgcggc
     2701 caacgccgga tcagggcgag ataactgggc aacaattgac tccagtgcgg gtctgccccc
     2761 tccgttggat gcaatagcca ctacctgctc aggggtggag acgaccggtg gaaagcgggc
     2821 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     2881 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     2941 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     3001 ctggaggacc tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc
     3061 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3121 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3181 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     3241 cgcccgtctc cttgagcggg gcacccgtca gtgcattgcg ccatgcatgc actgcctcca
     3301 ctgcggtcac gccgccacgt tttgcaatct tgagaagttg gcctgtgtcc aactgtaacg
     3361 gtggacctct caactctccc gccaccgtga gcaaggcctc cagagcgcgt gcgccggacc
     3421 actgtttgcc gacgccaacg atcgcttcgt gtgtcgcctc tggcaacgct gcgatcatgt
     3481 cctgatactt gacagcgacg gtccctaacg ctgccgggtg ttggcttaac gcaacgatgt
     3541 gcgcgtgtgt aaacccgtgg ccgaccagtg cctcgtggtg ctgcgccact gtcgaacgaa
     3601 ccttcggttt gatcttctcc tgttgctgct ggctgtagcc gagcgtgcgt agatcggcga
     3661 tatcgatgct gtccatgtgc tgtctctcga acttggcagc ggcggtctcc ttatcaactt
     3721 tattatacaa agttggcatt ataaaaaagc attgctcatc aatttgttgc aacgaacagg
     3781 tcactatcag tcaaaataaa atcattattt ggggcccgag cttaagactg gccgtcgttt
     3841 tacaacgtcg tgactgggaa aacatccatg ctagcgttaa cgcgagagta gggaactgcc
     3901 aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg cctttcgttt tatctgttgt
     3961 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgtg
     4021 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatc
//
