LOCUS       sequence-379081-        4075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD 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, 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 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: 
            "10.1038/s41477-021-00954-6"; journalName: "Nat Plants"; date: 
            "2021-07"; volume: "7"; issue: "7"; pages: "906-913"
COMMENT     Sequence Label: E3_pENTR_L3-L2_NN_G1333-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     377..394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(553..580)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(672..758)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     822..838
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    855..950
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             953..997
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1307..1717)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1582..1598
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1608..1626
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1674..1694)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1674..1692)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1713..1729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1726..1748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1737..1753
                     /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(1761..1767,1768..1785,1786..1791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1806..1827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2199..2450
                     /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(2472..2571)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2602..2620)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2625..2641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2743..2762)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2754..3563
                     /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(2821..2840)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3710..4075,1..223)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
       61 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
      121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
      181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      241 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
      301 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
      361 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
      421 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
      481 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt
      541 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
      601 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
      661 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      721 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      781 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct
      841 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
      901 aattgatgag caatgctttt ttataatgcc aactttgtat aataaagttg ataaggagac
      961 cgccgctgcc aagttcgaga gacagcacat ggacagcatc gatatcgccg atctacgcac
     1021 gctcggctac agccagcagc aacaggagaa gatcaaaccg aaggttcgtt cgacagtggc
     1081 gcagcaccac gaggcactgg tcggccacgg gtttacacac gcgcacatcg ttgcgttaag
     1141 ccaacacccg gcagcgttag ggaccgtcgc tgtcaagtat caggacatga tcgcagcgtt
     1201 gccagaggcg acacacgaag cgatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc
     1261 tctggaggcc ttgctcacgg tggcgggaga gttgagaggt ccaccgttac agttggacac
     1321 aggccaactt ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg cagtgcatgc
     1381 atggcgcaat gcactgacgg gtgccccgct caaggagacg ggcgccgcta cagggcgcgt
     1441 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     1501 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     1561 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     1621 tatagggcga attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag
     1681 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1741 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1801 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg
     1861 tcgtctccac ccctgagcag gtagtggcta ttgcatccaa caacgggggc agacccgcac
     1921 tggagtcaat tgttgcccag ttatctcgcc ctgatccggc gttggccgcg ttgaccaacg
     1981 accacctcgt cgccttggcc tgcctcggcg ggcgtcctgc gctggatgca gtgaaaaagg
     2041 gattgggggg atctggtagc tatgcgttag gaccctatca gatttcagct cctcaattgc
     2101 ctgcctataa tgggcaaact gttggcacct tttactacgt caatgatgct ggagggttag
     2161 aatccaaggt gttctcaagt ggtggttctg gaggtagtac gaatctttcg gacatcatag
     2221 agaaggaaac tggaaaacag ctcgttatcc aagagagcat tctcatgttg ccagaagaag
     2281 ttgaagaggt tataggcaac aaaccggaat ctgacattct ggtacatacc gcttatgatg
     2341 agtcaacaga tgagaacgtc atgcttttga catctgatgc accagaatac aaaccttggg
     2401 cacttgtgat tcaggattcc aatggtgaga acaagatcaa gatgctatga taactcgagc
     2461 gatcctctag aacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2521 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2581 tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     2641 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     2701 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2761 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     2821 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     2881 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2941 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     3001 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     3061 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     3121 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     3181 gcgtatttcg ccttgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     3241 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     3301 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     3361 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3421 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3481 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3541 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     3601 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     3661 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacac
//
