LOCUS       sequence-379196-        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_NI_G1397-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..57
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(216..243)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(335..421)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     485..501
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    517..616
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             619..645
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(952..1362)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1227..1243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1253..1271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1319..1339)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1319..1337)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1358..1374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1371..1393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1382..1398
                     /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(1406..1412,1413..1430,1431..1436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1451..1472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1802..2053
                     /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(2079..2174)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2200..2218)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2223..2239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2341..2360)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2352..3161
                     /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(2419..2438)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3308..3896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3797..3816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
       61 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
      181 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
      241 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
      301 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
      361 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
      421 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
      481 cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg
      541 atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg
      601 tacaaaaaag caggctttta cccatacgat gttccagatt acgctgatat cgccgatcta
      661 cgcacgctcg gctacagcca gcagcaacag gagaagatca aaccgaaggt tcgttcgaca
      721 gtggcgcagc accacgaggc actggtcggc cacgggttta cacacgcgca catcgttgcg
      781 ttaagccaac acccggcagc gttagggacc gtcgctgtca agtatcagga catgatcgca
      841 gcgttgccag aggcgacaca cgaagcgatc gttggcgtcg gcaaacagtg gtccggcgca
      901 cgcgctctgg aggccttgct cacggtggcg ggagagttga gaggtccacc gttacagttg
      961 gacacaggcc aacttctcaa gattgcaaaa cgtggcggcg tgaccgcagt ggaggcagtg
     1021 catgcatggc gcaatgcact gacgggtgcc ccgctcaagg agacgggcgc cgctacaggg
     1081 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1141 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1201 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     1261 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc ttttgttccc
     1321 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1501 accggtcgtc tccacccctg agcaggtagt ggctattgca tccaacatag ggggcagacc
     1561 cgcactggag tcaattgttg cccagttatc tcgccctgat ccggcgttgg ccgcgttgac
     1621 caacgaccac ctcgtcgcct tggcctgcct cggcgggcgt cctgcgctgg atgcagtgaa
     1681 aaagggattg gggggttctg cgattccagt taagagagga gctacaggag aaacgaaagt
     1741 ctttactggg aattccaatt ctcccaaatc accgattaaa ggcggatgta gtggtggtag
     1801 taccaatctt tccgacatta tcgagaagga aacaggtaaa caactcgtaa tccaagaaag
     1861 catactgatg cttcctgaag aggttgaaga ggtcataggg aacaaacctg aaagcgacat
     1921 tttggttcat actgcctatg atgagtctac agatgagaac gtgatgttgc taacctcaga
     1981 tgcacctgaa tacaagccat gggctttagt gattcaggat tcgaatggag agaacaagat
     2041 caagatgctc tgataactcg agcgatcctc tagacaccca acttttctat acaaagttgg
     2101 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2161 taaaatcatt atttgccatc cagctgcagg gcggccgccc ctatagtgag tcgtattaca
     2221 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     2281 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2341 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2401 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2461 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2521 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2581 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2641 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2701 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2761 gtccttttaa cagcgatcgc gtatttcgcc ttgctcaggc gcaatcacga atgaataacg
     2821 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2881 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2941 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3001 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3061 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3121 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     3181 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     3241 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3421 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3721 ccgaagggag aaaggcggac aggtatccgg taagcggcat ggtcggaaca ggagagcgca
     3781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3901 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3961 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
//
