LOCUS       sequence-379195-        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_NN_G1397-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    241..340
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             343..369
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(676..1086)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     951..967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        977..995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1043..1063)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1043..1061)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1082..1098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1095..1117)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1106..1122
                     /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(1130..1136,1137..1154,1155..1160))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1175..1196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1526..1777
                     /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(1803..1898)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1924..1942)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1947..1963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2065..2084)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2076..2885
                     /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(2143..2162)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3032..3620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3521..3540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3774..3791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3950..3977)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
      241 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
      301 tgctttttta taatgccaac tttgtacaaa aaagcaggct tttacccata cgatgttcca
      361 gattacgctg atatcgccga tctacgcacg ctcggctaca gccagcagca acaggagaag
      421 atcaaaccga aggttcgttc gacagtggcg cagcaccacg aggcactggt cggccacggg
      481 tttacacacg cgcacatcgt tgcgttaagc caacacccgg cagcgttagg gaccgtcgct
      541 gtcaagtatc aggacatgat cgcagcgttg ccagaggcga cacacgaagc gatcgttggc
      601 gtcggcaaac agtggtccgg cgcacgcgct ctggaggcct tgctcacggt ggcgggagag
      661 ttgagaggtc caccgttaca gttggacaca ggccaacttc tcaagattgc aaaacgtggc
      721 ggcgtgaccg cagtggaggc agtgcatgca tggcgcaatg cactgacggg tgccccgctc
      781 aaggagacgg gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
      841 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      901 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      961 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
     1021 cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     1081 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1141 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1201 cgttgcgctc actgcccgct ttccaccggt cgtctccacc cctgagcagg tagtggctat
     1261 tgcatccaac aacgggggca gacccgcact ggagtcaatt gttgcccagt tatctcgccc
     1321 tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct gcctcggcgg
     1381 gcgtcctgcg ctggatgcag tgaaaaaggg attggggggt tctgcgattc cagttaagag
     1441 aggagctaca ggagaaacga aagtctttac tgggaattcc aattctccca aatcaccgat
     1501 taaaggcgga tgtagtggtg gtagtaccaa tctttccgac attatcgaga aggaaacagg
     1561 taaacaactc gtaatccaag aaagcatact gatgcttcct gaagaggttg aagaggtcat
     1621 agggaacaaa cctgaaagcg acattttggt tcatactgcc tatgatgagt ctacagatga
     1681 gaacgtgatg ttgctaacct cagatgcacc tgaatacaag ccatgggctt tagtgattca
     1741 ggattcgaat ggagagaaca agatcaagat gctctgataa ctcgagcgat cctctagaca
     1801 cccaactttt ctatacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1861 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc
     1921 gcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     1981 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     2041 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     2101 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     2161 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     2221 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2281 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2341 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     2401 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2461 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgccttgctc
     2521 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2581 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2641 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2701 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2761 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2821 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2881 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     2941 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     3001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3481 gcatggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3721 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3781 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3841 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3901 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     3961 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
//
