LOCUS       sequence-379080-        4075 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 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_NI_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(931..1740)
                     /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     1654..1673
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1732..1751
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1853..1869
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1874..1892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1923..2022
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2044..2295)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    2667..2688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2703..2708,2709..2726,2727..2733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2741..2757
                     /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     2746..2768
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2765..2781
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2777..3187
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     2800..2820
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2802..2820
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2868..2886)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2896..2912)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3497..3541)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    complement(3544..3639)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3656..3672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3736..3822
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3914..3941
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact
      841 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
      901 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
      961 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1021 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1081 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1141 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     1201 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1261 atcaaccaaa ccgttattca ttcgtgattg cgcctgagca aggcgaaata cgcgatcgct
     1321 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1381 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc
     1441 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1501 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1561 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1621 gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1681 atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat
     1741 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     1801 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac
     1861 agctatgacc atgtaatacg actcactata ggggatatcg cggccgccct gcagctggat
     1921 ggcaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa attgataagc
     1981 aatgctttct tataatgcca actttgtaca agaaagctgg gttctagagg atcgctcgag
     2041 ttatcatagc atcttgatct tgttctcacc attggaatcc tgaatcacaa gtgcccaagg
     2101 tttgtattct ggtgcatcag atgtcaaaag catgacgttc tcatctgttg actcatcata
     2161 agcggtatgt accagaatgt cagattccgg tttgttgcct ataacctctt caacttcttc
     2221 tggcaacatg agaatgctct cttggataac gagctgtttt ccagtttcct tctctatgat
     2281 gtccgaaaga ttcgtactac ctccagaacc accacttgag aacaccttgg attctaaccc
     2341 tccagcatca ttgacgtagt aaaaggtgcc aacagtttgc ccattatagg caggcaattg
     2401 aggagctgaa atctgatagg gtcctaacgc atagctacca gatcccccca atcccttttt
     2461 cactgcatcc agcgcaggac gcccgccgag gcaggccaag gcgacgaggt ggtcgttggt
     2521 caacgcggcc aacgccggat cagggcgaga taactgggca acaattgact ccagtgcggg
     2581 tctgccccct atgttggatg caatagccac tacctgctca ggggtggaga cgaccggtgg
     2641 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2701 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2761 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     2821 gaacaaaagc tggaggacct cgaggggggg cccggtaccc aattcgccct atagtgagtc
     2881 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2941 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3001 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     3061 ctgtagcggc gcccgtctcc ttgagcgggg cacccgtcag tgcattgcgc catgcatgca
     3121 ctgcctccac tgcggtcacg ccgccacgtt ttgcaatctt gagaagttgg cctgtgtcca
     3181 actgtaacgg tggacctctc aactctcccg ccaccgtgag caaggcctcc agagcgcgtg
     3241 cgccggacca ctgtttgccg acgccaacga tcgcttcgtg tgtcgcctct ggcaacgctg
     3301 cgatcatgtc ctgatacttg acagcgacgg tccctaacgc tgccgggtgt tggcttaacg
     3361 caacgatgtg cgcgtgtgta aacccgtggc cgaccagtgc ctcgtggtgc tgcgccactg
     3421 tcgaacgaac cttcggtttg atcttctcct gttgctgctg gctgtagccg agcgtgcgta
     3481 gatcggcgat atcgatgctg tccatgtgct gtctctcgaa cttggcagcg gcggtctcct
     3541 tatcaacttt attatacaaa gttggcatta taaaaaagca ttgctcatca atttgttgca
     3601 acgaacaggt cactatcagt caaaataaaa tcattatttg gggcccgagc ttaagactgg
     3661 ccgtcgtttt acaacgtcgt gactgggaaa acatccatgc tagcgttaac gcgagagtag
     3721 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt
     3781 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     3841 aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     3901 catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     3961 cctggctagc ggtacgcgta ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4021 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggttt
//
