LOCUS       sequence-379227-        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_NG_G1397-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /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="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     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             2147..2398
                     /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(2424..2519)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2545..2563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2568..2584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2686..2705)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2697..3506
                     /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(2764..2783)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3653..4010,1..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 tcgaggtcct 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 ttgcatccaa cggagggggc agacccgcac tggagtcaat
     1921 tgttgcccag ttatctcgcc ctgatccggc gttggccgcg ttgaccaacg accacctcgt
     1981 cgccttggcc tgcctcggcg ggcgtcctgc gctggatgca gtgaaaaagg gattgggggg
     2041 ttctgcgatt ccagttaaga gaggagctac aggagaaacg aaagtcttta ctgggaattc
     2101 caattctccc aaatcaccga ctaaaggcgg atgtagtggt ggtagtacca atctttccga
     2161 cattatcgag aaggaaacag gtaaacaact cgtaatccaa gaaagcatac tgatgcttcc
     2221 tgaagaggtt gaagaggtca tagggaacaa acctgaaagc gacattttgg ttcatactgc
     2281 ctatgatgag tctacagatg agaacgtgat gttgctaacc tcagatgcac ctgaatacaa
     2341 gccatgggct ttagtgattc aggattcgaa tggagagaac aagatcaaga tgctctgata
     2401 actcgagcga tcctctagac acccaacttt tctatacaaa gttggcatta taagaaagca
     2461 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2521 ccatccagct gcagggcggc cgcccctata gtgagtcgta ttacatggtc atagctgttt
     2581 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2641 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     2701 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     2761 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     2821 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2881 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     2941 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     3001 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     3061 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     3121 atcgcgtatt tcgccttgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     3181 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     3241 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     3301 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     3361 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     3421 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     3481 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     3541 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     3601 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3781 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3841 ctgtagcacc gcctacatac cttgctctgc taatcctgtt accagtggct gctgccagtg
     3901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
//
