LOCUS       sequence-393343-        4202 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 4202)
  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 4202)
  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_G1333-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(9..95)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     159..175
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    191..290
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             293..319
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(626..1036)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     901..917
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        927..945
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(993..1013)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(993..1011)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1032..1048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1045..1067)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1056..1072
                     /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(1080..1086,1087..1104,1105..1110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1125..1146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1668..1919
                     /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(1945..2040)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2066..2084)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2089..2105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2207..2226)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2218..3027
                     /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(2285..2304)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3174..3762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3663..3682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3916..3933
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4092..4119)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
       61 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
      121 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
      181 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
      241 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct tttacccata
      301 cgatgttcca gattacgctg atatcgccga tctacgcacg ctcggctaca gccagcagca
      361 acaggagaag atcaaaccga aggttcgttc gacagtggcg cagcaccacg aggcactggt
      421 cggccacggg tttacacacg cgcacatcgt tgcgttaagc caacacccgg cagcgttagg
      481 gaccgtcgct gtcaagtatc aggacatgat cgcagcgttg ccagaggcga cacacgaagc
      541 gatcgttggc gtcggcaaac agtggtccgg cgcacgcgct ctggaggcct tgctcacggt
      601 ggcgggagag ttgagaggtc caccgttaca gttggacaca ggccaacttc tcaagattgc
      661 aaaacgtggc ggcgtgaccg cagtggaggc agtgcatgca tggcgcaatg cactgacggg
      721 tgccccgctc aaggagacgg gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
      781 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      841 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      901 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
      961 ggccccccct cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1021 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1081 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1141 acattaattg cgttgcgctc actgcccgct ttccaccggt cgtctccacc cctgagcagg
     1201 tagtggctat tgcatccaac atagggggca gacccgcact ggagtcaatt gttgcccagt
     1261 tatctcgccc tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct
     1321 gcctcggcgg gcgtcctgcg ctggatgcag tgaaaaaggg attggggggt agtccaactc
     1381 catatcccaa ttatgtgagt gcaggccatg ttgaaggtca atcagcccta ttcatgagag
     1441 ataacggaat aagtgaagga ctggtgtttc acaacaatcc aaaaggtact tgtggatttt
     1501 gcgtaaacat gattgaaacc cttctcccag aaaacgctaa gatgacagta gttccacctg
     1561 aaggtgctat tcctgtcaaa agaggtgcta ctggtgaaac caaggtgttt actgggaatt
     1621 ccaattcacc caaaagccca atcaaaggtg ggtgtagtgg aggatctaca aatctctctg
     1681 acatcattga gaaagagact ggaaagcaac tagtcattca ggagtcaatc ctgatgttac
     1741 cagaggaggt tgaggaagtg ataggcaata agcccgaaag cgatatactt gttcatactg
     1801 cctatgacga atcgacggat gagaacgtaa tgcttctaac ctcagatgct cctgagtaca
     1861 aaccttgggc gttagttatc caggattcca atggagagaa caagatcaag atgttgtgat
     1921 aactcgagcg atcctctaga cacccaactt ttctatacaa agttggcatt ataagaaagc
     1981 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     2041 gccatccagc tgcagggcgg ccgcccctat agtgagtcgt attacatggt catagctgtt
     2101 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     2161 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2221 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     2281 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     2341 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2401 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2461 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     2521 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2581 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2641 gatcgcgtat ttcgccttgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2701 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2761 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2821 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2881 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2941 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     3001 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     3061 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     3121 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3601 gcggacaggt atccggtaag cggcatggtc ggaacaggag agcgcacgag ggagcttcca
     3661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3841 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3901 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3961 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4021 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     4081 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     4141 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     4201 ct
//
