LOCUS       sequence-379225-        4244 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 4244)
  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 4244)
  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_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..85
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(244..271)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(363..449)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     513..529
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    545..644
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             647..673
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(980..1390)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1255..1271
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1281..1299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1347..1367)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1347..1365)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1386..1402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1399..1421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1410..1426
                     /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(1434..1440,1441..1458,1459..1464))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1479..1500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2064..2315
                     /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(2341..2436)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2462..2480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2485..2501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2603..2622)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2614..3423
                     /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(2681..2700)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3570..4158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4059..4078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
       61 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      121 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      181 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
      241 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
      301 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
      361 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
      421 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac
      481 gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg
      541 gccccaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgatga
      601 gcaatgcttt tttataatgc caactttgta caaaaaagca ggcttttacc catacgatgt
      661 tccagattac gctgatatcg ccgatctacg cacgctcggc tacagccagc agcaacagga
      721 gaagatcaaa ccgaaggttc gttcgacagt ggcgcagcac cacgaggcac tggtcggcca
      781 cgggtttaca cacgcgcaca tcgttgcgtt aagccaacac ccggcagcgt tagggaccgt
      841 cgctgtcaag tatcaggaca tgatcgcagc gttgccagag gcgacacacg aagcgatcgt
      901 tggcgtcggc aaacagtggt ccggcgcacg cgctctggag gccttgctca cggtggcggg
      961 agagttgaga ggtccaccgt tacagttgga cacaggccaa cttctcaaga ttgcaaaacg
     1021 tggcggcgtg accgcagtgg aggcagtgca tgcatggcgc aatgcactga cgggtgcccc
     1081 gctcaaggag acgggcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     1141 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     1201 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     1261 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
     1321 ccctcgaggc cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     1381 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1441 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1501 attgcgttgc gctcactgcc cgctttccac cggtcgtctc cacccctgag caggtagtgg
     1561 ctattgcatc ccacgacggg ggcagacccg cactggagtc aattgttgcc cagttatctc
     1621 gccctgatcc ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg
     1681 gcgggcgtcc tgcgctggat gcagtgaaaa agggattggg ggggtctgga tcgtatgctt
     1741 taggaccgta tcagatctca gctccacaat tgcctgcata taacggacaa actgttggga
     1801 ccttttacta cgttaacgat gctggtggat tggagtccaa agtgttctct tctggtggcc
     1861 caactccata tcccaattat gcgaatgcag gccatgttga aggtcaatca gccctattca
     1921 tgagagataa cggaataagt gaaggactgg tgtttcacaa caatccagaa ggtacttgtg
     1981 gattttgcgt aaacatgact gagacacttc tcccagaaaa tgccaagatg acagttgtac
     2041 ctcctgaagg ttctggtgga tcgacaaacc tttcagacat tatcgagaaa gagacaggca
     2101 aacagctagt gattcaagag tccattctca tgcttcccga agaagttgag gaagtcattg
     2161 ggaataagcc ggaaagtgac atactcgttc atacggctta cgatgagagc acggatgaga
     2221 atgtcatgtt gcttaccagt gatgcacctg aatacaaacc ttgggcttta gtcatccagg
     2281 acagcaatgg tgagaacaag atcaagatgc tgtgataact cgagcgatcc tctagacacc
     2341 caacttttct atacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2401 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gggcggccgc
     2461 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2521 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2581 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2641 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2701 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2761 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2821 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2881 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2941 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     3001 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg ccttgctcag
     3061 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     3121 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     3181 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     3241 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3301 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3361 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3421 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3481 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3541 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3601 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3661 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3721 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3781 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3841 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3901 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3961 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4021 atggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4081 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4141 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4201 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attc
//
