LOCUS       sequence-379198-        4010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD_v2 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, Itoh T, Tsutsumi N, Arimura SI
  TITLE     Characterization and development of a plastid genome base editor, 
            ptpTALECD.
  JOURNAL   Plant J. 2023 Aug;115(4):1151-1162. doi: 10.1111/tpj.16311. Epub 
            2023 Jun 2.
  PUBMED    37265080
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: "10.1111/tpj.16311";
            journalName: "Plant J"; date: "2023-08"; volume: "115"; issue: "4"; 
            pages: "1151-1162"
COMMENT     Sequence Label: E1_pENTR_L1-L4_HD_G1397-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..46
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(205..232)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(324..410)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     474..490
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    506..605
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             608..634
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(941..1351)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1216..1232
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1242..1260
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1308..1328)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1308..1326)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1347..1363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1360..1382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1371..1387
                     /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(1395..1401,1402..1419,1420..1425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1440..1461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1791..2042
                     /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(2068..2163)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2189..2207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2212..2228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2330..2349)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2341..3150
                     /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(2408..2427)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3297..3885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3786..3805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
       61 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      121 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
      181 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
      241 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
      301 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      361 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
      421 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
      481 gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg
      541 ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc
      601 aggcttttac ccatacgatg ttccagatta cgctgatatc gccgatctac gcacgctcgg
      661 ctacagccag cagcaacagg agaagatcaa accgaaggtt cgttcgacag tggcgcagca
      721 ccacgaggca ctggtcggcc acgggtttac acacgcgcac atcgttgcgt taagccaaca
      781 cccggcagcg ttagggaccg tcgctgtcaa gtatcaggac atgatcgcag cgttgccaga
      841 ggcgacacac gaagcgatcg ttggcgtcgg caaacagtgg tccggcgcac gcgctctgga
      901 ggccttgctc acggtggcgg gagagttgag aggtccaccg ttacagttgg acacaggcca
      961 acttctcaag attgcaaaac gtggcggcgt gaccgcagtg gaggcagtgc atgcatggcg
     1021 caatgcactg acgggtgccc cgctcaagga gacgggcgcc gctacagggc gcgtcccatt
     1081 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     1141 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     1201 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     1261 gcgaattggg taccgggccc cccctcgagg ccctccagct tttgttccct ttagtgaggg
     1321 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1381 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1441 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtcgtct
     1501 ccacccctga gcaggtagtg gctattgcat cccacgacgg gggcagaccc gcactggagt
     1561 caattgttgc ccagttatct cgccctgatc cggcgttggc cgcgttgacc aacgaccacc
     1621 tcgtcgcctt ggcctgcctc ggcgggcgtc ctgcgctgga tgcagtgaaa aagggattgg
     1681 ggggttctgc gattccagtt aagagaggag ctacaggaga aacgaaagtc tttactggga
     1741 attccaattc tcccaaatca ccgattaaag gcggatgtag tggtggtagt accaatcttt
     1801 ccgacattat cgagaaggaa acaggtaaac aactcgtaat ccaagaaagc atactgatgc
     1861 ttcctgaaga ggttgaagag gtcataggga acaaacctga aagcgacatt ttggttcata
     1921 ctgcctatga tgagtctaca gatgagaacg tgatgttgct aacctcagat gcacctgaat
     1981 acaagccatg ggctttagtg attcaggatt cgaatggaga gaacaagatc aagatgctct
     2041 gataactcga gcgatcctct agacacccaa cttttctata caaagttggc attataagaa
     2101 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     2161 tttgccatcc agctgcaggg cggccgcccc tatagtgagt cgtattacat ggtcatagct
     2221 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2281 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2341 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2401 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2461 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2521 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2581 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2641 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2701 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2761 agcgatcgcg tatttcgcct tgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2821 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2881 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2941 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     3001 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     3061 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3121 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3181 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3241 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3301 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3361 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3421 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3481 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3541 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3601 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3661 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3721 aaggcggaca ggtatccggt aagcggcatg gtcggaacag gagagcgcac gagggagctt
     3781 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3841 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3901 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3961 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
//
