LOCUS       sequence-379072-        4225 bp    DNA     circular SYN 15-APR-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 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 Apr 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-C
FEATURES             Location/Qualifiers
     source          1..4225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..87)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(241..829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(321..340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(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     1699..1718
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1777..1796
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1898..1914
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1919..1937
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1968..2067
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2089..2340)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    2862..2883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2898..2903,2904..2921,2922..2928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2936..2952
                     /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     2941..2963
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2960..2976
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2972..3382
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     2995..3015
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2997..3015
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3063..3081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3091..3107)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3692..3736)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    complement(3739..3834)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3851..3867)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3931..4017
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4109..4136
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
       61 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      841 gatcttttct acggggtctg acgctcagtg gaacgacgcg taactcacgt taagggattt
      901 tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca
      961 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1021 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1081 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1141 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1201 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     1261 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     1321 attcattcgt gattgcgcct gagcaaggcg aaatacgcga tcgctgttaa aaggacaatt
     1381 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1441 acctgaatca ggatattctt ctaatacctg gaatgctgtt tttccgggga tcgcagtggt
     1501 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1561 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1621 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc
     1681 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1741 ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     1801 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     1861 gtaacatcag agattttgag acacgggcca gagctgccag gaaacagcta tgaccatgta
     1921 atacgactca ctatagggga tatcgcggcc gccctgcagc tggatggcaa ataatgattt
     1981 tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc tttcttataa
     2041 tgccaacttt gtacaagaaa gctgggttct agaggatcgc tcgagttatc acaacatctt
     2101 gatcttgttc tctccattgg aatcctggat aactaacgcc caaggtttgt actcaggagc
     2161 atctgaggtt agaagcatta cgttctcatc cgtcgattcg tcataggcag tatgaacaag
     2221 tatatcgctt tcgggcttat tgcctatcac ttcctcaacc tcctctggta acatcaggat
     2281 tgactcctga atgactagtt gctttccagt ctctttctca atgatgtcag agagatttgt
     2341 agatcctcca ctacacccac ctttcgttgg gcttttgggt gaattggaat tcccagtaaa
     2401 caccttggtt tcaccagtag cacctctttt gacaggaata gcaccttcag gtggaactac
     2461 tgtcatctta gcgttttctg ggagaagggt ttctgtcatg ttgacgcaaa atccacatgt
     2521 tccttcaggg ttgttgtgga acaccaaccc ttcagaaatg ccgttatccc tcatgaacaa
     2581 tgcagattga ccttcaacat gtcctgcatt ggcgtaattc ggatacggag ttggactacc
     2641 ccccaatccc tttttcactg catccagcgc aggacgcccg ccgaggcagg ccaaggcgac
     2701 gaggtggtcg ttggtcaacg cggccaacgc cggatcaggg cgagataact gggcaacaat
     2761 tgactccagt gcgggtctgc ccccgttgtt ggatgcaata gccactacct gctcaggggt
     2821 ggagacgacc ggtggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2881 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2941 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     3001 aaccctcact aaagggaaca aaagctggag gacctcgagg gggggcccgg tacccaattc
     3061 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     3121 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3181 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3241 cgaatgggac gcgccctgta gcggcgcccg tctccttgag cggggcaccc gtcagtgcat
     3301 tgcgccatgc atgcactgcc tccactgcgg tcacgccgcc acgttttgca atcttgagaa
     3361 gttggcctgt gtccaactgt aacggtggac ctctcaactc tcccgccacc gtgagcaagg
     3421 cctccagagc gcgtgcgccg gaccactgtt tgccgacgcc aacgatcgct tcgtgtgtcg
     3481 cctctggcaa cgctgcgatc atgtcctgat acttgacagc gacggtccct aacgctgccg
     3541 ggtgttggct taacgcaacg atgtgcgcgt gtgtaaaccc gtggccgacc agtgcctcgt
     3601 ggtgctgcgc cactgtcgaa cgaaccttcg gtttgatctt ctcctgttgc tgctggctgt
     3661 agccgagcgt gcgtagatcg gcgatatcga tgctgtccat gtgctgtctc tcgaacttgg
     3721 cagcggcggt ctccttatca actttattat acaaagttgg cattataaaa aagcattgct
     3781 catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggggcc
     3841 cgagcttaag actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg
     3901 ttaacgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac
     3961 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4021 ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4081 ccataaactg ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg
     4141 tttctacaaa ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg
     4201 ctttccagtc gggaaacctg tcgtg
//
