LOCUS       sequence-393339-        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_HD_G1333-DddA11'-C
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      69..96
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(255..272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(426..1014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(506..525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1161..1970)
                     /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     1884..1903
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1962..1981
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2083..2099
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2104..2122
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2148..2243
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2269..2520)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    3042..3063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3078..3083,3084..3101,3102..3108)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3116..3132
                     /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     3121..3143
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3140..3156
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3152..3562
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRASRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3175..3195
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3177..3195
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3243..3261)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3271..3287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3869..3895)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3898..3997)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4013..4029)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4093..4179
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
       61 aactaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttcctgg
      121 ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      181 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      241 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      301 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      361 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      421 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      481 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      541 gtgcgctctc ctgttccgac catgccgctt accggatacc tgtccgcctt tctcccttcg
      601 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      661 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      721 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      781 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      841 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      901 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      961 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1021 cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact cacgttaagg
     1081 gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg ccagtgttac
     1141 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     1201 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     1261 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1321 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1381 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     1441 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1501 ccgttattca ttcgtgattg cgcctgagca aggcgaaata cgcgatcgct gttaaaagga
     1561 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1621 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc ggggatcgca
     1681 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1741 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1801 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt
     1861 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1921 atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat aacacccctt
     1981 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     2041 gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac agctatgacc
     2101 atgtaatacg actcactata ggggcggccg ccctgcagct ggatggcaaa taatgatttt
     2161 attttgactg atagtgacct gttcgttgca acaaattgat aagcaatgct ttcttataat
     2221 gccaactttg tatagaaaag ttgggtgtct agaggatcgc tcgagttatc acaacatctt
     2281 gatcttgttc tctccattgg aatcctggat aactaacgcc caaggtttgt actcaggagc
     2341 atctgaggtt agaagcatta cgttctcatc cgtcgattcg tcataggcag tatgaacaag
     2401 tatatcgctt tcgggcttat tgcctatcac ttcctcaacc tcctctggta acatcaggat
     2461 tgactcctga atgactagtt gctttccagt ctctttctca atgatgtcag agagatttgt
     2521 agatcctcca ctacacccac ctttgattgg gcttttgggt gaattggaat tcccagtaaa
     2581 caccttggtt tcaccagtag cacctctttt gacaggaata gcaccttcag gtggaactac
     2641 tgtcatctta gcgttttctg ggagaagggt ttcaatcatg tttacgcaaa atccacaagt
     2701 accttttgga ttgttgtgaa acaccagtcc ttcacttatt ccgttatctc tcatgaatag
     2761 ggctgattga ccttcaacat ggcctgcact cacataattg ggatatggag ttggactacc
     2821 ccccaatccc tttttcactg catccagcgc aggacgcccg ccgaggcagg ccaaggcgac
     2881 gaggtggtcg ttggtcaacg cggccaacgc cggatcaggg cgagataact gggcaacaat
     2941 tgactccagt gcgggtctgc ccccgtcgtg ggatgcaata gccactacct gctcaggggt
     3001 ggagacgacc ggtggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     3061 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     3121 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     3181 aaccctcact aaagggaaca aaagctggag ggcctcgagg gggggcccgg tacccaattc
     3241 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     3301 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3361 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3421 cgaatgggac gcgccctgta gcggcgcccg tctccttgag cggggcaccc gtcagtgcat
     3481 tgcgccatgc atgcactgcc tccactgcgg tcacgccgcc acgttttgca atcttgagaa
     3541 gttggcctgt gtccaactgt aacggtggac ctctcaactc tcccgccacc gtgagcaagg
     3601 cctccagagc gcgtgcgccg gaccactgtt tgccgacgcc aacgatcgct tcgtgtgtcg
     3661 cctctggcaa cgctgcgatc atgtcctgat acttgacagc gacggtccct aacgctgccg
     3721 ggtgttggct taacgcaacg atgtgcgcgt gtgtaaaccc gtggccgacc agtgcctcgt
     3781 ggtgctgcgc cactgtcgaa cgaaccttcg gtttgatctt ctcctgttgc tgctggctgt
     3841 agccgagcgt gcgtagatcg gcgatatcag cgtaatctgg aacatcgtat gggtaaaagc
     3901 ctgctttttt gtacaaagtt ggcattataa aaaagcattg ctcatcaatt tgttgcaacg
     3961 aacaggtcac tatcagtcaa aataaaatca ttatttgggg cccgagctta agactggccg
     4021 tcgttttaca acgtcgtgac tgggaaaaca tccatgctag cgttaacgcg agagtaggga
     4081 actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc
     4141 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     4201 gt
//
