LOCUS       sequence-379108-        4267 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 4267)
  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 4267)
  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_G1397-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(54..140)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     204..220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    237..332
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             335..379
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(689..1099)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     964..980
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        990..1008
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1056..1076)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1056..1074)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1095..1111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1108..1130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1119..1135
                     /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(1143..1149,1150..1167,1168..1173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1188..1209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1773..2024
                     /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(2046..2145)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2176..2194)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2199..2215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2317..2336)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2328..3137
                     /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(2395..2414)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3284..3872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3773..3792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4026..4043
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4202..4229)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
       61 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
      121 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
      181 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat
      241 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt
      301 ttttataatg ccaactttgt ataataaagt tgataaggag accgccgctg ccaagttcga
      361 gagacagcac atggacagca tcgatatcgc cgatctacgc acgctcggct acagccagca
      421 gcaacaggag aagatcaaac cgaaggttcg ttcgacagtg gcgcagcacc acgaggcact
      481 ggtcggccac gggtttacac acgcgcacat cgttgcgtta agccaacacc cggcagcgtt
      541 agggaccgtc gctgtcaagt atcaggacat gatcgcagcg ttgccagagg cgacacacga
      601 agcgatcgtt ggcgtcggca aacagtggtc cggcgcacgc gctctggagg ccttgctcac
      661 ggtggcggga gagttgagag gtccaccgtt acagttggac acaggccaac ttctcaagat
      721 tgcaaaacgt ggcggcgtga ccgcagtgga ggcagtgcat gcatggcgca atgcactgac
      781 gggtgccccg ctcaaggaga cgggcgccgc tacagggcgc gtcccattcg ccattcaggc
      841 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      901 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      961 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     1021 ccgggccccc cctcgaggtc ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1201 ctcacattaa ttgcgttgcg ctcactgccc gctttccacc ggtcgtctcc acccctgagc
     1261 aggtagtggc tattgcatcc aacatagggg gcagacccgc actggagtca attgttgccc
     1321 agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc gtcgccttgg
     1381 cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg gggtctggat
     1441 cgtatgcttt aggaccgtat cagatctcag ctccacaatt gcctgcatat aacggacaaa
     1501 ctgttgggac cttttactac gttaacgatg ctggtggatt ggagtccaaa gtgttcatct
     1561 ctggtggccc aactccatat cccaattatg tgagtgcagg ccatgttgaa ggtcaatcag
     1621 ccctattcat gagagataac ggaataagtg aaggactggt gtttcacaac aatccaaaag
     1681 gtacttgtgg attttgcgta aacatgattg agacacttct cccagaaaat gccaagatga
     1741 cagttgtacc tcctgaaggt tctggtggat cgacaaacct ttcagacatt atcgagaaag
     1801 agacaggcaa acagctagtg attcaagagt ccattctcat gcttcccgaa gaagttgagg
     1861 aagtcattgg gaataagccg gaaagtgaca tactcgttca tacggcttac gatgagagca
     1921 cggatgagaa tgtcatgttg cttaccagtg atgcacctga atacaaacct tgggctttag
     1981 tcatccagga cagcaatggt gagaacaaga tcaagatgct gtgataactc gagcgatcct
     2041 ctagaaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2101 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagg
     2161 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2221 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2281 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2341 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2401 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     2461 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2521 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2581 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2641 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2701 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2761 ttcgccttgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2821 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     2881 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2941 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3001 aggatcttgc cattctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3061 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3121 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3181 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3301 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3361 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3421 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3481 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3541 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3601 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3661 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3721 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3781 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3841 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3901 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3961 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4021 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4081 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4141 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4201 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     4261 gcgtcct
//
