LOCUS       sequence-376585-        4202 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 4202)
  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 4202)
  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-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     163..182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     416..433
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(592..619)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(711..797)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     861..877
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    893..992
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             995..1021
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1328..1738)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1603..1619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1629..1647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1695..1715)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1695..1713)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1734..1750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1747..1769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1758..1774
                     /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(1782..1788,1789..1806,1807..1812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1827..1848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2370..2621
                     /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(2647..2742)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2768..2786)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2791..2807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2909..2928)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2920..3729
                     /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(2987..3006)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3876..4202,1..262)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
       61 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      121 cggcatggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      301 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      361 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
      421 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      481 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      541 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
      601 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
      661 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
      721 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
      781 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
      841 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
      901 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
      961 tataatgcca actttgtaca aaaaagcagg cttttaccca tacgatgttc cagattacgc
     1021 tgatatcgcc gatctacgca cgctcggcta cagccagcag caacaggaga agatcaaacc
     1081 gaaggttcgt tcgacagtgg cgcagcacca cgaggcactg gtcggccacg ggtttacaca
     1141 cgcgcacatc gttgcgttaa gccaacaccc ggcagcgtta gggaccgtcg ctgtcaagta
     1201 tcaggacatg atcgcagcgt tgccagaggc gacacacgaa gcgatcgttg gcgtcggcaa
     1261 acagtggtcc ggcgcacgcg ctctggaggc cttgctcacg gtggcgggag agttgagagg
     1321 tccaccgtta cagttggaca caggccaact tctcaagatt gcaaaacgtg gcggcgtgac
     1381 cgcagtggag gcagtgcatg catggcgcaa tgcactgacg ggtgccccgc tcaaggagac
     1441 gggcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     1501 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1561 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     1621 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcc
     1681 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     1741 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1801 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1861 tcactgcccg ctttccaccg gtcgtctcca cccctgagca ggtagtggct attgcatcca
     1921 acggaggggg cagacccgca ctggagtcaa ttgttgccca gttatctcgc cctgatccgg
     1981 cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc gggcgtcctg
     2041 cgctggatgc agtgaaaaag ggattggggg gtagtccaac tccgtatccg aattacgcca
     2101 atgcaggaca tgttgaaggt caatctgcat tgttcatgag ggataacggc atttctgaag
     2161 ggttggtgtt ccacaacaac cctgaaggaa catgtggatt ttgcgtcaac atgacagaaa
     2221 cccttctccc agaaaacgct aagatgacag tagttccacc tgaaggtgct attcctgtca
     2281 aaagaggtgc tactggtgaa accaaggtgt ttactgggaa ttccaattca cccaaaagcc
     2341 caacgaaagg tgggtgtagt ggaggatcta caaatctctc tgacatcatt gagaaagaga
     2401 ctggaaagca actagtcatt caggagtcaa tcctgatgtt accagaggag gttgaggaag
     2461 tgataggcaa taagcccgaa agcgatatac ttgttcatac tgcctatgac gaatcgacgg
     2521 atgagaacgt aatgcttcta acctcagatg ctcctgagta caaaccttgg gcgttagtta
     2581 tccaggattc caatggagag aacaagatca agatgttgtg ataactcgag cgatcctcta
     2641 gacacccaac ttttctatac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2701 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagggc
     2761 ggccgcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2821 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2881 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2941 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     3001 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     3061 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     3121 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     3181 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3241 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3301 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctt
     3361 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3421 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3481 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3541 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3601 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3661 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3721 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     3781 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3841 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4201 gg
//
