LOCUS       sequence-393342-        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_NN_G1333-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      84..111
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(270..287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(441..1029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(521..540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1176..1985)
                     /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     1899..1918
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1977..1996
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2098..2114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2119..2137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2163..2258
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2284..2535)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    3057..3078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3093..3098,3099..3116,3117..3123)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3131..3147
                     /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)."
     primer_bind     3136..3158
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3155..3171
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3167..3577
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3190..3210
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3192..3210
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3258..3276)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3286..3302)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3884..3910)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3913..4012)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4028..4044)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4108..4194
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcggatttg aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata
       61 aactgccagg catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct
      121 acaaactctt cctggctagc ggtacgcgta ttaattgcgt tgcgctcact gcccgctttc
      181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccatgc cgcttaccgg atacctgtcc
      601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
     1081 taactcacgt taagggattt tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg
     1141 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     1201 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1261 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1321 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1381 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1441 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1501 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcaaggcg aaatacgcga
     1561 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1621 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1681 tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1741 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1801 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1861 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1921 tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg
     1981 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     2041 atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgccag
     2101 gaaacagcta tgaccatgta atacgactca ctataggggc ggccgccctg cagctggatg
     2161 gcaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca
     2221 atgctttctt ataatgccaa ctttgtatag aaaagttggg tgtctagagg atcgctcgag
     2281 ttatcacaac atcttgatct tgttctctcc attggaatcc tggataacta acgcccaagg
     2341 tttgtactca ggagcatctg aggttagaag cattacgttc tcatccgtcg attcgtcata
     2401 ggcagtatga acaagtatat cgctttcggg cttattgcct atcacttcct caacctcctc
     2461 tggtaacatc aggattgact cctgaatgac tagttgcttt ccagtctctt tctcaatgat
     2521 gtcagagaga tttgtagatc ctccactaca cccacctttg attgggcttt tgggtgaatt
     2581 ggaattccca gtaaacacct tggtttcacc agtagcacct cttttgacag gaatagcacc
     2641 ttcaggtgga actactgtca tcttagcgtt ttctgggaga agggtttcaa tcatgtttac
     2701 gcaaaatcca caagtacctt ttggattgtt gtgaaacacc agtccttcac ttattccgtt
     2761 atctctcatg aatagggctg attgaccttc aacatggcct gcactcacat aattgggata
     2821 tggagttgga ctacccccca atcccttttt cactgcatcc agcgcaggac gcccgccgag
     2881 gcaggccaag gcgacgaggt ggtcgttggt caacgcggcc aacgccggat cagggcgaga
     2941 taactgggca acaattgact ccagtgcggg tctgcccccg ttgttggatg caatagccac
     3001 tacctgctca ggggtggaga cgaccggtgg aaagcgggca gtgagcgcaa cgcaattaat
     3061 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     3121 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     3181 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggaggacct cgaggggggg
     3241 cccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     3301 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3361 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3421 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcccgtctcc ttgagcgggg
     3481 cacccgtcag tgcattgcgc catgcatgca ctgcctccac tgcggtcacg ccgccacgtt
     3541 ttgcaatctt gagaagttgg cctgtgtcca actgtaacgg tggacctctc aactctcccg
     3601 ccaccgtgag caaggcctcc agagcgcgtg cgccggacca ctgtttgccg acgccaacga
     3661 tcgcttcgtg tgtcgcctct ggcaacgctg cgatcatgtc ctgatacttg acagcgacgg
     3721 tccctaacgc tgccgggtgt tggcttaacg caacgatgtg cgcgtgtgta aacccgtggc
     3781 cgaccagtgc ctcgtggtgc tgcgccactg tcgaacgaac cttcggtttg atcttctcct
     3841 gttgctgctg gctgtagccg agcgtgcgta gatcggcgat atcagcgtaa tctggaacat
     3901 cgtatgggta aaagcctgct tttttgtaca aagttggcat tataaaaaag cattgctcat
     3961 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tggggcccga
     4021 gcttaagact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat gctagcgtta
     4081 acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg
     4141 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4201 gg
//
