LOCUS       sequence-338618-        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_NI_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     269..288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     522..539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(698..725)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(817..903)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     967..983
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    999..1098
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1101..1127
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1434..1844)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1709..1725
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1735..1753
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1801..1821)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1801..1819)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1840..1856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1853..1875)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1864..1880
                     /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(1888..1894,1895..1912,1913..1918))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1933..1954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2518..2769
                     /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(2795..2890)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2916..2934)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2939..2955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3057..3076)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3068..3877
                     /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(3135..3154)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(4024..4244,1..368)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
       61 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      121 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      181 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc atggtcggaa
      241 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      301 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      361 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      421 ctcacatgtt ctttcctgcg ttatcccccg attctgtgga taaccgtatt accgcctttg
      481 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      541 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      601 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
      661 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
      721 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
      781 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
      841 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
      901 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
      961 gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac
     1021 tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt
     1081 tgtacaaaaa agcaggcttt tacccatacg atgttccaga ttacgctgat atcgccgatc
     1141 tacgcacgct cggctacagc cagcagcaac aggagaagat caaaccgaag gttcgttcga
     1201 cagtggcgca gcaccacgag gcactggtcg gccacgggtt tacacacgcg cacatcgttg
     1261 cgttaagcca acacccggca gcgttaggga ccgtcgctgt caagtatcag gacatgatcg
     1321 cagcgttgcc agaggcgaca cacgaagcga tcgttggcgt cggcaaacag tggtccggcg
     1381 cacgcgctct ggaggccttg ctcacggtgg cgggagagtt gagaggtcca ccgttacagt
     1441 tggacacagg ccaacttctc aagattgcaa aacgtggcgg cgtgaccgca gtggaggcag
     1501 tgcatgcatg gcgcaatgca ctgacgggtg ccccgctcaa ggagacgggc gccgctacag
     1561 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     1621 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     1681 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     1741 gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc
     1801 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1861 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1921 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1981 ccaccggtcg tctccacccc tgagcaggta gtggctattg catccaacat agggggcaga
     2041 cccgcactgg agtcaattgt tgcccagtta tctcgccctg atccggcgtt ggccgcgttg
     2101 accaacgacc acctcgtcgc cttggcctgc ctcggcgggc gtcctgcgct ggatgcagtg
     2161 aaaaagggat tgggggggtc tggatcgtat gctttaggac cgtatcagat ctcagctcca
     2221 caattgcctg catataacgg acaaactgtt gggacctttt actacgttaa cgatgctggt
     2281 ggattggagt ccaaagtgtt ctcttctggt ggcccaactc catatcccaa ttatgcgaat
     2341 gcaggccatg ttgaaggtca atcagcccta ttcatgagag ataacggaat aagtgaagga
     2401 ctggtgtttc acaacaatcc agaaggtact tgtggatttt gcgtaaacat gactgagaca
     2461 cttctcccag aaaatgccaa gatgacagtt gtacctcctg aaggttctgg tggatcgaca
     2521 aacctttcag acattatcga gaaagagaca ggcaaacagc tagtgattca agagtccatt
     2581 ctcatgcttc ccgaagaagt tgaggaagtc attgggaata agccggaaag tgacatactc
     2641 gttcatacgg cttacgatga gagcacggat gagaatgtca tgttgcttac cagtgatgca
     2701 cctgaataca aaccttgggc tttagtcatc caggacagca atggtgagaa caagatcaag
     2761 atgctgtgat aactcgagcg atcctctaga cacccaactt ttctatacaa agttggcatt
     2821 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     2881 atcattattt gccatccagc tgcagggcgg ccgcccctat agtgagtcgt attacatggt
     2941 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     3001 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     3061 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     3121 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     3181 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     3241 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     3301 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     3361 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     3421 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     3481 ttttaacagc gatcgcgtat ttcgccttgc tcaggcgcaa tcacgaatga ataacggttt
     3541 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3601 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3661 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3721 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3781 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3841 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3901 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3961 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4021 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4081 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4141 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4201 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaa
//
