LOCUS       sequence-379231-        4052 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 4052)
  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 4052)
  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_NG_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             965..991
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1298..1708)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1573..1589
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1599..1617
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1665..1685)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1665..1683)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1704..1720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1717..1739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1728..1744
                     /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(1752..1758,1759..1776,1777..1782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1797..1818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2190..2441
                     /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(2467..2562)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2588..2606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2611..2627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2729..2748)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2740..3549
                     /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(2807..2826)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3696..4052,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcatggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
      961 cttttaccca tacgatgttc cagattacgc tgatatcgcc gatctacgca cgctcggcta
     1021 cagccagcag caacaggaga agatcaaacc gaaggttcgt tcgacagtgg cgcagcacca
     1081 cgaggcactg gtcggccacg ggtttacaca cgcgcacatc gttgcgttaa gccaacaccc
     1141 ggcagcgtta gggaccgtcg ctgtcaagta tcaggacatg atcgcagcgt tgccagaggc
     1201 gacacacgaa gcgatcgttg gcgtcggcaa acagtggtcc ggcgcacgcg ctctggaggc
     1261 cttgctcacg gtggcgggag agttgagagg tccaccgtta cagttggaca caggccaact
     1321 tctcaagatt gcaaaacgtg gcggcgtgac cgcagtggag gcagtgcatg catggcgcaa
     1381 tgcactgacg ggtgccccgc tcaaggagac gggcgccgct acagggcgcg tcccattcgc
     1441 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     1501 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     1561 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     1621 aattgggtac cgggcccccc ctcgaggtcc tccagctttt gttcccttta gtgagggtta
     1681 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1741 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1801 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtcgtctcca
     1861 cccctgagca ggtagtggct attgcatcca acggaggggg cagacccgca ctggagtcaa
     1921 ttgttgccca gttatctcgc cctgatccgg cgttggccgc gttgaccaac gaccacctcg
     1981 tcgccttggc ctgcctcggc gggcgtcctg cgctggatgc agtgaaaaag ggattggggg
     2041 gatctggtag ctatgcgtta ggaccctatc agatttcagc tcctcaattg cctgcctata
     2101 atgggcaaac tgttggcacc ttttactacg tcaatgatgc tggagggtta gaatccaagg
     2161 tgttctcaag tggtggttct ggaggtagta cgaatctttc ggacatcata gagaaggaaa
     2221 ctggaaaaca gctcgttatc caagagagca ttctcatgtt gccagaagaa gttgaagagg
     2281 ttataggcaa caaaccggaa tctgacattc tggtacatac cgcttatgat gagtcaacag
     2341 atgagaacgt catgcttttg acatctgatg caccagaata caaaccttgg gcacttgtga
     2401 ttcaggattc caatggtgag aacaagatca agatgctatg ataactcgag cgatcctcta
     2461 gacacccaac ttttctatac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2521 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagggc
     2581 ggccgcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2641 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2701 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2761 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2821 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2881 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2941 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     3001 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3061 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3121 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctt
     3181 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3241 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3301 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3361 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3421 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3481 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3541 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     3601 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3841 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4021 ggttcgtgca cacagcccag cttggagcga ac
//
