LOCUS       sequence-393336-        4075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD_v2 vector assembly, Entry clone 3 with attL2 and L3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4075)
  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 4075)
  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: E3_pENTR_L3-L2_NG_G1333-DddA11'-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     414..431
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(590..617)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(709..795)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     859..875
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    892..987
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             990..1034
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1344..1754)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1619..1635
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1645..1663
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1711..1731)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1711..1729)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1750..1766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1763..1785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1774..1790
                     /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(1798..1804,1805..1822,1823..1828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1843..1864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2236..2487
                     /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(2509..2608)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2639..2657)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2662..2678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2780..2799)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2791..3600
                     /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(2858..2877)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3747..4075,1..260)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
       61 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      301 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      361 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
      421 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      481 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      541 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
      601 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
      661 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
      721 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
      781 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
      841 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga
      901 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta
      961 taatgccaac tttgtataat aaagttgata aggagaccgc cgctgccaag ttcgagagac
     1021 agcacatgga cagcatcgat atcgccgatc tacgcacgct cggctacagc cagcagcaac
     1081 aggagaagat caaaccgaag gttcgttcga cagtggcgca gcaccacgag gcactggtcg
     1141 gccacgggtt tacacacgcg cacatcgttg cgttaagcca acacccggca gcgttaggga
     1201 ccgtcgctgt caagtatcag gacatgatcg cagcgttgcc agaggcgaca cacgaagcga
     1261 tcgttggcgt cggcaaacag tggtccggcg cacgcgctct ggaggccttg ctcacggtgg
     1321 cgggagagtt gagaggtcca ccgttacagt tggacacagg ccaacttctc aagattgcaa
     1381 aacgtggcgg cgtgaccgca gtggaggcag tgcatgcatg gcgcaatgca ctgacgggtg
     1441 ccccgctcaa ggagacgggc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     1501 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     1561 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     1621 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg
     1681 ccccccctcg aggtcctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     1741 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1801 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1861 attaattgcg ttgcgctcac tgcccgcttt ccaccggtcg tctccacccc tgagcaggta
     1921 gtggctattg catccaacgg agggggcaga cccgcactgg agtcaattgt tgcccagtta
     1981 tctcgccctg atccggcgtt ggccgcgttg accaacgacc acctcgtcgc cttggcctgc
     2041 ctcggcgggc gtcctgcgct ggatgcagtg aaaaagggat tggggggatc tggtagctat
     2101 gcgttaggac cctatcagat ttcagctcct caattgcctg cctataatgg gcaaactgtt
     2161 ggcacctttt actacgtcaa tgatgctgga gggttagaat ccaaggtgtt cataagtggt
     2221 ggttctggag gtagtacgaa tctttcggac atcatagaga aggaaactgg aaaacagctc
     2281 gttatccaag agagcattct catgttgcca gaagaagttg aagaggttat aggcaacaaa
     2341 ccggaatctg acattctggt acataccgct tatgatgagt caacagatga gaacgtcatg
     2401 cttttgacat ctgatgcacc agaatacaaa ccttgggcac ttgtgattca ggattccaat
     2461 ggtgagaaca agatcaagat gctatgataa ctcgagcgat cctctagaac ccagctttct
     2521 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     2581 ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg cgatatcccc
     2641 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     2701 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     2761 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     2821 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2881 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2941 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     3001 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     3061 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     3121 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     3181 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgcct tgctcaggcg
     3241 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     3301 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     3361 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3421 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     3481 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     3541 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     3601 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     3661 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     3721 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3781 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3841 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3901 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3961 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4021 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggtt
//
