LOCUS       sequence-338458-        4010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 1 with attL1 and L4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4010)
  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 4010)
  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: E1_pENTR_L1-L4_NN_G1397-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(197..785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(277..296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(932..1741)
                     /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     1655..1674
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1733..1752
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1854..1870
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1875..1893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1919..2014
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2040..2291)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2731..3141
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2822..2840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2850..2866)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3448..3474)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3477..3576)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3592..3608)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3672..3758
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3850..3877
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
       61 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      121 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      181 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      301 gaagctccct cgtgcgctct cctgttccga ccatgccgct taccggatac ctgtccgcct
      361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgtaac
      841 tcacgttaag ggattttggt catgagcttg cgccgtcccg tcaagtcagc gtaatgctct
      901 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
      961 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1021 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1081 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1141 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     1201 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1261 catcaaccaa accgttattc attcgtgatt gcgcctgagc aaggcgaaat acgcgatcgc
     1321 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     1381 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgtttttc
     1441 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1501 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1561 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1621 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1681 aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca
     1741 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     1801 ttttatcttg tgcaatgtaa catcagagat tttgagacac gggccagagc tgccaggaaa
     1861 cagctatgac catgtaatac gactcactat aggggcggcc gccctgcagc tggatggcaa
     1921 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
     1981 tttcttataa tgccaacttt gtatagaaaa gttgggtgtc tagaggatcg ctcgagttat
     2041 cagagcatct tgatcttgtt ctctccattc gaatcctgaa tcactaaagc ccatggcttg
     2101 tattcaggtg catctgaggt tagcaacatc acgttctcat ctgtagactc atcataggca
     2161 gtatgaacca aaatgtcgct ttcaggtttg ttccctatga cctcttcaac ctcttcagga
     2221 agcatcagta tgctttcttg gattacgagt tgtttacctg tttccttctc gataatgtcg
     2281 gaaagattgg tactaccacc actacatccg cctttagtcg gtgatttggg agaattggaa
     2341 ttcccagtaa agactttcgt ttctcctgta gctcctctct taactggaat cgcagaaccc
     2401 cccaatccct ttttcactgc atccagcgca ggacgcccgc cgaggcaggc caaggcgacg
     2461 aggtggtcgt tggtcaacgc ggccaacgcc ggatcagggc gagataactg ggcaacaatt
     2521 gactccagtg cgggtctgcc cccgttgttg gatgcaatag ccactacctg ctcaggggtg
     2581 gagacgaccg gtggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctggagg acctcgaggg ggggcccggt acccaattcg
     2821 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     2881 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2941 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3001 gaatgggacg cgccctgtag cggcgcccgt ctccttgagc ggggcacccg tcagtgcatt
     3061 gcgccatgca tgcactgcct ccactgcggt cacgccgcca cgttttgcaa tcttgagaag
     3121 ttggcctgtg tccaactgta acggtggacc tctcaactct cccgccaccg tgagcaaggc
     3181 ctccagagcg cgtgcgccgg accactgttt gccgacgcca acgatcgctt cgtgtgtcgc
     3241 ctctggcaac gctgcgatca tgtcctgata cttgacagcg acggtcccta acgctgccgg
     3301 gtgttggctt aacgcaacga tgtgcgcgtg tgtaaacccg tggccgacca gtgcctcgtg
     3361 gtgctgcgcc actgtcgaac gaaccttcgg tttgatcttc tcctgttgct gctggctgta
     3421 gccgagcgtg cgtagatcgg cgatatcagc gtaatctgga acatcgtatg ggtaaaagcc
     3481 tgcttttttg tacaaagttg gcattataaa aaagcattgc tcatcaattt gttgcaacga
     3541 acaggtcact atcagtcaaa ataaaatcat tatttggggc ccgagcttaa gactggccgt
     3601 cgttttacaa cgtcgtgact gggaaaacat ccatgctagc gttaacgcga gagtagggaa
     3661 ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga ctgggccttt cgttttatct
     3721 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3781 ttgtgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3841 aaactaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttcctg
     3901 gctagcggta cgcgtattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
//
