LOCUS       sequence-379110-        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_HD_G1397-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(69..155)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     219..235
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    252..347
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             350..394
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(704..1114)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     979..995
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1005..1023
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1071..1091)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1071..1089)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1110..1126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1123..1145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1134..1150
                     /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(1158..1164,1165..1182,1183..1188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1203..1224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1788..2039
                     /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(2061..2160)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2191..2209)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2214..2230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2332..2351)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2343..3152
                     /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(2410..2429)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3299..3887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3788..3807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4041..4058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4217..4244)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
       61 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
      121 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
      181 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
      241 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
      301 tgatgagcaa tgctttttta taatgccaac tttgtataat aaagttgata aggagaccgc
      361 cgctgccaag ttcgagagac agcacatgga cagcatcgat atcgccgatc tacgcacgct
      421 cggctacagc cagcagcaac aggagaagat caaaccgaag gttcgttcga cagtggcgca
      481 gcaccacgag gcactggtcg gccacgggtt tacacacgcg cacatcgttg cgttaagcca
      541 acacccggca gcgttaggga ccgtcgctgt caagtatcag gacatgatcg cagcgttgcc
      601 agaggcgaca cacgaagcga tcgttggcgt cggcaaacag tggtccggcg cacgcgctct
      661 ggaggccttg ctcacggtgg cgggagagtt gagaggtcca ccgttacagt tggacacagg
      721 ccaacttctc aagattgcaa aacgtggcgg cgtgaccgca gtggaggcag tgcatgcatg
      781 gcgcaatgca ctgacgggtg ccccgctcaa ggagacgggc gccgctacag ggcgcgtccc
      841 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      901 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      961 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     1021 agggcgaatt gggtaccggg ccccccctcg aggccctcca gcttttgttc cctttagtga
     1081 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1141 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1201 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccaccggtcg
     1261 tctccacccc tgagcaggta gtggctattg catcccacga cgggggcaga cccgcactgg
     1321 agtcaattgt tgcccagtta tctcgccctg atccggcgtt ggccgcgttg accaacgacc
     1381 acctcgtcgc cttggcctgc ctcggcgggc gtcctgcgct ggatgcagtg aaaaagggat
     1441 tgggggggtc tggatcgtat gctttaggac cgtatcagat ctcagctcca caattgcctg
     1501 catataacgg acaaactgtt gggacctttt actacgttaa cgatgctggt ggattggagt
     1561 ccaaagtgtt catctctggt ggcccaactc catatcccaa ttatgtgagt gcaggccatg
     1621 ttgaaggtca atcagcccta ttcatgagag ataacggaat aagtgaagga ctggtgtttc
     1681 acaacaatcc aaaaggtact tgtggatttt gcgtaaacat gattgagaca cttctcccag
     1741 aaaatgccaa gatgacagtt gtacctcctg aaggttctgg tggatcgaca aacctttcag
     1801 acattatcga gaaagagaca ggcaaacagc tagtgattca agagtccatt ctcatgcttc
     1861 ccgaagaagt tgaggaagtc attgggaata agccggaaag tgacatactc gttcatacgg
     1921 cttacgatga gagcacggat gagaatgtca tgttgcttac cagtgatgca cctgaataca
     1981 aaccttgggc tttagtcatc caggacagca atggtgagaa caagatcaag atgctgtgat
     2041 aactcgagcg atcctctaga acccagcttt cttgtacaaa gttggcatta taagaaagca
     2101 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2161 ccatccagct gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
     2221 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2281 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2341 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2401 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2461 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2521 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2581 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2641 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2701 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2761 acagcgatcg cgtatttcgc cttgctcagg cgcaatcacg aatgaataac ggtttggttg
     2821 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2881 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2941 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3001 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3061 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3121 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3181 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3241 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3301 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3361 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3421 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3481 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3541 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3601 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3661 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3721 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3781 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3841 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3901 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3961 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4021 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4081 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4141 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     4201 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     4261 ggcagtt
//
