LOCUS       sequence-379229-        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_HD_G1397-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     366..383
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(542..569)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(661..747)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     811..827
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    843..942
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             945..971
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1278..1688)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1553..1569
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1579..1597
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1645..1665)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1645..1663)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1684..1700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1697..1719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1708..1724
                     /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(1732..1738,1739..1756,1757..1762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1777..1798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2128..2379
                     /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(2405..2500)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2526..2544)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2549..2565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2667..2686)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2678..3487
                     /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(2745..2764)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3634..4010,1..212)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
       61 atccggtaag cggcatggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      301 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      361 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      481 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
      541 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
      601 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
      661 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
      721 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      781 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      841 cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc
      901 aatgcttttt tataatgcca actttgtaca aaaaagcagg cttttaccca tacgatgttc
      961 cagattacgc tgatatcgcc gatctacgca cgctcggcta cagccagcag caacaggaga
     1021 agatcaaacc gaaggttcgt tcgacagtgg cgcagcacca cgaggcactg gtcggccacg
     1081 ggtttacaca cgcgcacatc gttgcgttaa gccaacaccc ggcagcgtta gggaccgtcg
     1141 ctgtcaagta tcaggacatg atcgcagcgt tgccagaggc gacacacgaa gcgatcgttg
     1201 gcgtcggcaa acagtggtcc ggcgcacgcg ctctggaggc cttgctcacg gtggcgggag
     1261 agttgagagg tccaccgtta cagttggaca caggccaact tctcaagatt gcaaaacgtg
     1321 gcggcgtgac cgcagtggag gcagtgcatg catggcgcaa tgcactgacg ggtgccccgc
     1381 tcaaggagac gggcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     1441 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     1501 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     1561 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     1621 ctcgaggccc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     1681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1801 tgcgttgcgc tcactgcccg ctttccaccg gtcgtctcca cccctgagca ggtagtggct
     1861 attgcatccc acgacggggg cagacccgca ctggagtcaa ttgttgccca gttatctcgc
     1921 cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc ctgcctcggc
     1981 gggcgtcctg cgctggatgc agtgaaaaag ggattggggg gttctgcgat tccagttaag
     2041 agaggagcta caggagaaac gaaagtcttt actgggaatt ccaattctcc caaatcaccg
     2101 actaaaggcg gatgtagtgg tggtagtacc aatctttccg acattatcga gaaggaaaca
     2161 ggtaaacaac tcgtaatcca agaaagcata ctgatgcttc ctgaagaggt tgaagaggtc
     2221 atagggaaca aacctgaaag cgacattttg gttcatactg cctatgatga gtctacagat
     2281 gagaacgtga tgttgctaac ctcagatgca cctgaataca agccatgggc tttagtgatt
     2341 caggattcga atggagagaa caagatcaag atgctctgat aactcgagcg atcctctaga
     2401 cacccaactt ttctatacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     2461 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgcagggcgg
     2521 ccgcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2581 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2641 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2701 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2761 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2821 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2881 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2941 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3001 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3061 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgccttgc
     3121 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3181 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3241 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3301 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3361 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3421 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3481 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3541 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3781 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
//
