LOCUS       sequence-393335-        4075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD_v2 vector assembly, Entry clone 3 with attL2 and L3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4075)
  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 4075)
  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: E3_pENTR_L3-L2_NI_G1333-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      81..108
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(267..284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(438..1026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(518..537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1173..1982)
                     /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     1896..1915
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1974..1993
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2095..2111
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2116..2134
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2165..2264
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2286..2537)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    2909..2930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2945..2950,2951..2968,2969..2975)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2983..2999
                     /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)."
     primer_bind     2988..3010
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3007..3023
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3019..3429
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3042..3062
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3044..3062
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3110..3128)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3138..3154)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3739..3783)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    complement(3786..3881)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3898..3914)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3978..4064
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggatttgaac gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
       61 tgccaggcat caaactaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
      121 aactcttcct ggctagcggt acgcgtatta attgcgttgc gctcactgcc cgctttccag
      181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgtaa
     1081 ctcacgttaa gggattttgg tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc
     1141 tgccagtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga
     1201 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     1261 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     1321 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     1381 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     1441 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     1501 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag caaggcgaaa tacgcgatcg
     1561 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     1621 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt
     1681 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     1741 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     1801 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     1861 aagcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     1921 aaatcagcat ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc
     1981 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     2041 tttttatctt gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgccaggaa
     2101 acagctatga ccatgtaata cgactcacta taggggatat cgcggccgcc ctgcagctgg
     2161 atggcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa
     2221 gcaatgcttt cttataatgc caactttgta caagaaagct gggttctaga ggatcgctcg
     2281 agttatcata gcatcttgat cttgttctca ccattggaat cctgaatcac aagtgcccaa
     2341 ggtttgtatt ctggtgcatc agatgtcaaa agcatgacgt tctcatctgt tgactcatca
     2401 taagcggtat gtaccagaat gtcagattcc ggtttgttgc ctataacctc ttcaacttct
     2461 tctggcaaca tgagaatgct ctcttggata acgagctgtt ttccagtttc cttctctatg
     2521 atgtccgaaa gattcgtact acctccagaa ccaccactta tgaacacctt ggattctaac
     2581 cctccagcat cattgacgta gtaaaaggtg ccaacagttt gcccattata ggcaggcaat
     2641 tgaggagctg aaatctgata gggtcctaac gcatagctac cagatccccc caatcccttt
     2701 ttcactgcat ccagcgcagg acgcccgccg aggcaggcca aggcgacgag gtggtcgttg
     2761 gtcaacgcgg ccaacgccgg atcagggcga gataactggg caacaattga ctccagtgcg
     2821 ggtctgcccc ctatgttgga tgcaatagcc actacctgct caggggtgga gacgaccggt
     2881 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     2941 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     3001 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     3061 gggaacaaaa gctggaggac ctcgaggggg ggcccggtac ccaattcgcc ctatagtgag
     3121 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3181 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3241 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     3301 ccctgtagcg gcgcccgtct ccttgagcgg ggcacccgtc agtgcattgc gccatgcatg
     3361 cactgcctcc actgcggtca cgccgccacg ttttgcaatc ttgagaagtt ggcctgtgtc
     3421 caactgtaac ggtggacctc tcaactctcc cgccaccgtg agcaaggcct ccagagcgcg
     3481 tgcgccggac cactgtttgc cgacgccaac gatcgcttcg tgtgtcgcct ctggcaacgc
     3541 tgcgatcatg tcctgatact tgacagcgac ggtccctaac gctgccgggt gttggcttaa
     3601 cgcaacgatg tgcgcgtgtg taaacccgtg gccgaccagt gcctcgtggt gctgcgccac
     3661 tgtcgaacga accttcggtt tgatcttctc ctgttgctgc tggctgtagc cgagcgtgcg
     3721 tagatcggcg atatcgatgc tgtccatgtg ctgtctctcg aacttggcag cggcggtctc
     3781 cttatcaact ttattataca aagttggcat tataaaaaag cattgctcat caatttgttg
     3841 caacgaacag gtcactatca gtcaaaataa aatcattatt tggggcccga gcttaagact
     3901 ggccgtcgtt ttacaacgtc gtgactggga aaacatccat gctagcgtta acgcgagagt
     3961 agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt
     4021 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagc
//
