LOCUS       sequence-379197-        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_NG_G1397-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(163..190)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(282..368)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     432..448
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    464..563
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             566..592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(899..1309)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1174..1190
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1200..1218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1266..1286)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1266..1284)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1305..1321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1318..1340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1329..1345
                     /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(1353..1359,1360..1377,1378..1383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1398..1419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1749..2000
                     /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(2026..2121)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2147..2165)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2170..2186)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2288..2307)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2299..3108
                     /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(2366..2385)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3255..3843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3744..3763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3997..4010,1..4)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
       61 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      121 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
      181 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
      241 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
      301 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
      361 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
      421 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt
      481 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc
      541 aactttgtac aaaaaagcag gcttttaccc atacgatgtt ccagattacg ctgatatcgc
      601 cgatctacgc acgctcggct acagccagca gcaacaggag aagatcaaac cgaaggttcg
      661 ttcgacagtg gcgcagcacc acgaggcact ggtcggccac gggtttacac acgcgcacat
      721 cgttgcgtta agccaacacc cggcagcgtt agggaccgtc gctgtcaagt atcaggacat
      781 gatcgcagcg ttgccagagg cgacacacga agcgatcgtt ggcgtcggca aacagtggtc
      841 cggcgcacgc gctctggagg ccttgctcac ggtggcggga gagttgagag gtccaccgtt
      901 acagttggac acaggccaac ttctcaagat tgcaaaacgt ggcggcgtga ccgcagtgga
      961 ggcagtgcat gcatggcgca atgcactgac gggtgccccg ctcaaggaga cgggcgccgc
     1021 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1081 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1141 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     1201 aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc ctccagcttt
     1261 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     1321 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1381 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1441 gctttccacc ggtcgtctcc acccctgagc aggtagtggc tattgcatcc aacggagggg
     1501 gcagacccgc actggagtca attgttgccc agttatctcg ccctgatccg gcgttggccg
     1561 cgttgaccaa cgaccacctc gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg
     1621 cagtgaaaaa gggattgggg ggttctgcga ttccagttaa gagaggagct acaggagaaa
     1681 cgaaagtctt tactgggaat tccaattctc ccaaatcacc gattaaaggc ggatgtagtg
     1741 gtggtagtac caatctttcc gacattatcg agaaggaaac aggtaaacaa ctcgtaatcc
     1801 aagaaagcat actgatgctt cctgaagagg ttgaagaggt catagggaac aaacctgaaa
     1861 gcgacatttt ggttcatact gcctatgatg agtctacaga tgagaacgtg atgttgctaa
     1921 cctcagatgc acctgaatac aagccatggg ctttagtgat tcaggattcg aatggagaga
     1981 acaagatcaa gatgctctga taactcgagc gatcctctag acacccaact tttctataca
     2041 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     2101 gtcaaaataa aatcattatt tgccatccag ctgcagggcg gccgccccta tagtgagtcg
     2161 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     2221 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     2281 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     2341 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2401 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2461 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2521 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2581 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     2641 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2701 tgtaattgtc cttttaacag cgatcgcgta tttcgccttg ctcaggcgca atcacgaatg
     2761 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2821 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     2881 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2941 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3001 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3061 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     3121 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     3181 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     3241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3361 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat accttgctct gctaatcctg
     3481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcatggt cggaacagga
     3721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3901 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3961 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
//
