LOCUS       sequence-338625-        4225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 1 with attL2 and L3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4225)
  AUTHORS   Nakazato I, Okuno M, Yamamoto H, Tamura Y, Itoh T, Shikanai T, 
            Takanashi H, Tsutsumi N, Arimura SI
  TITLE     Targeted base editing in the plastid genome of Arabidopsis thaliana.
  JOURNAL   Nat Plants. 2021 Jul;7(7):906-913. doi: 10.1038/s41477-021-00954-6. 
            Epub 2021 Jul 1.
  PUBMED    34211131
REFERENCE   2  (bases 1 to 4225)
  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: 
            "10.1038/s41477-021-00954-6"; journalName: "Nat Plants"; date: 
            "2021-07"; volume: "7"; issue: "7"; pages: "906-913"
COMMENT     Sequence Label: E3_pENTR_L3-L2_NG_G1333-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             421..672
                     /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(694..793)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(824..842)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(847..863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(965..984)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             976..1785
                     /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(1043..1062)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1932..2520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2421..2440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2674..2691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2850..2877)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2969..3055)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3119..3135
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3152..3247
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3250..3294
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(3604..4014)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3879..3895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3905..3923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3971..3991)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3971..3989)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4010..4026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4023..4045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4034..4050
                     /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(4058..4064,4065..4082,4083..4088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
       61 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
      121 ggtagtccaa ctccgtatcc gaattacgcc aatgcaggac atgttgaagg tcaatctgca
      181 ttgttcatga gggataacgg catttctgaa gggttggtgt tccacaacaa ccctgaagga
      241 acatgtggat tttgcgtcaa catgacagaa acccttctcc cagaaaacgc taagatgaca
      301 gtagttccac ctgaaggtgc tattcctgtc aaaagaggtg ctactggtga aaccaaggtg
      361 tttactggga attccaattc acccaaaagc ccaacgaaag gtgggtgtag tggaggatct
      421 acaaatctct ctgacatcat tgagaaagag actggaaagc aactagtcat tcaggagtca
      481 atcctgatgt taccagagga ggttgaggaa gtgataggca ataagcccga aagcgatata
      541 cttgttcata ctgcctatga cgaatcgacg gatgagaacg taatgcttct aacctcagat
      601 gctcctgagt acaaaccttg ggcgttagtt atccaggatt ccaatggaga gaacaagatc
      661 aagatgttgt gataactcga gcgatcctct agaacccagc tttcttgtac aaagttggca
      721 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
      781 aaatcattat ttgccatcca gctgcagggc ggccgcgata tcccctatag tgagtcgtat
      841 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
      901 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
      961 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     1021 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1081 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1141 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1201 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1261 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1321 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     1381 aattgtcctt ttaacagcga tcgcgtattt cgccttgctc aggcgcaatc acgaatgaat
     1441 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     1501 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     1561 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     1621 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     1681 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     1741 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     1801 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     1861 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     1921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2221 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2581 ttctttcctg cattatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2641 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2701 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2761 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     2821 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     2881 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     2941 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3001 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3061 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     3121 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg
     3181 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtataat
     3241 aaagttgata aggagaccgc cgctgccaag ttcgagagac agcacatgga cagcatcgat
     3301 atcgccgatc tacgcacgct cggctacagc cagcagcaac aggagaagat caaaccgaag
     3361 gttcgttcga cagtggcgca gcaccacgag gcactggtcg gccacgggtt tacacacgcg
     3421 cacatcgttg cgttaagcca acacccggca gcgttaggga ccgtcgctgt caagtatcag
     3481 gacatgatcg cagcgttgcc agaggcgaca cacgaagcga tcgttggcgt cggcaaacag
     3541 tggtccggcg cacgcgctct ggaggccttg ctcacggtgg cgggagagtt gagaggtcca
     3601 ccgttacagt tggacacagg ccaacttctc aagattgcaa aacgtggcgg cgtgaccgca
     3661 gtggaggcag tgcatgcatg gcgcaatgca ctgacgggtg ccccgctcaa ggagacgggc
     3721 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3781 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     3841 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
     3901 cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcctcca
     3961 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4141 tgcccgcttt ccaccggtcg tctccacccc tgagcaggta gtggctattg catccaacgg
     4201 agggggcaga cccgcactgg agtca
//
