LOCUS       sequence-376583-        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_HD_G1333-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(561..588)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(680..766)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     830..846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    862..961
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             964..990
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1297..1707)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1572..1588
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1598..1616
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1664..1684)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1664..1682)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1703..1719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1716..1738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1727..1743
                     /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(1751..1757,1758..1775,1776..1781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1796..1817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2339..2590
                     /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(2616..2711)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2737..2755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2760..2776)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2878..2897)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2889..3698
                     /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(2956..2975)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3845..4202,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcatggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
      541 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
      601 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
      661 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
      721 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
      781 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
      841 gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
      901 ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa aaaagcaggc
      961 ttttacccat acgatgttcc agattacgct gatatcgccg atctacgcac gctcggctac
     1021 agccagcagc aacaggagaa gatcaaaccg aaggttcgtt cgacagtggc gcagcaccac
     1081 gaggcactgg tcggccacgg gtttacacac gcgcacatcg ttgcgttaag ccaacacccg
     1141 gcagcgttag ggaccgtcgc tgtcaagtat caggacatga tcgcagcgtt gccagaggcg
     1201 acacacgaag cgatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc tctggaggcc
     1261 ttgctcacgg tggcgggaga gttgagaggt ccaccgttac agttggacac aggccaactt
     1321 ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc atggcgcaat
     1381 gcactgacgg gtgccccgct caaggagacg ggcgccgcta cagggcgcgt cccattcgcc
     1441 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     1501 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     1561 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     1621 attgggtacc gggccccccc tcgaggccct ccagcttttg ttccctttag tgagggttaa
     1681 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1741 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1801 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg tcgtctccac
     1861 ccctgagcag gtagtggcta ttgcatccca cgacgggggc agacccgcac tggagtcaat
     1921 tgttgcccag ttatctcgcc ctgatccggc gttggccgcg ttgaccaacg accacctcgt
     1981 cgccttggcc tgcctcggcg ggcgtcctgc gctggatgca gtgaaaaagg gattgggggg
     2041 tagtccaact ccgtatccga attacgccaa tgcaggacat gttgaaggtc aatctgcatt
     2101 gttcatgagg gataacggca tttctgaagg gttggtgttc cacaacaacc ctgaaggaac
     2161 atgtggattt tgcgtcaaca tgacagaaac ccttctccca gaaaacgcta agatgacagt
     2221 agttccacct gaaggtgcta ttcctgtcaa aagaggtgct actggtgaaa ccaaggtgtt
     2281 tactgggaat tccaattcac ccaaaagccc aacgaaaggt gggtgtagtg gaggatctac
     2341 aaatctctct gacatcattg agaaagagac tggaaagcaa ctagtcattc aggagtcaat
     2401 cctgatgtta ccagaggagg ttgaggaagt gataggcaat aagcccgaaa gcgatatact
     2461 tgttcatact gcctatgacg aatcgacgga tgagaacgta atgcttctaa cctcagatgc
     2521 tcctgagtac aaaccttggg cgttagttat ccaggattcc aatggagaga acaagatcaa
     2581 gatgttgtga taactcgagc gatcctctag acacccaact tttctataca aagttggcat
     2641 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2701 aatcattatt tgccatccag ctgcagggcg gccgccccta tagtgagtcg tattacatgg
     2761 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2821 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2881 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2941 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     3001 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     3061 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     3121 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     3181 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     3241 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     3301 cttttaacag cgatcgcgta tttcgccttg ctcaggcgca atcacgaatg aataacggtt
     3361 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     3421 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     3481 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     3541 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     3601 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     3661 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     3721 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3781 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3841 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3901 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3961 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4021 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4081 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4141 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4201 cg
//
