LOCUS       sequence-379218-        4033 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 4033)
  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 4033)
  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_G1397-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..4033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(561..588)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(680..766)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     830..846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..958
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             961..1005
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(1315..1725)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     1590..1606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1616..1634
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1682..1702)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1682..1700)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1721..1737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1734..1756)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1745..1761
                     /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(1769..1775,1776..1793,1794..1799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1814..1835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2165..2416
                     /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(2438..2537)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2568..2586)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2591..2607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2709..2728)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2720..3529
                     /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(2787..2806)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3676..4033,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
      541 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
      601 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
      661 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
      721 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
      781 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
      841 gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
      901 ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtataa taaagttgat
      961 aaggagaccg ccgctgccaa gttcgagaga cagcacatgg acagcatcga tatcgccgat
     1021 ctacgcacgc tcggctacag ccagcagcaa caggagaaga tcaaaccgaa ggttcgttcg
     1081 acagtggcgc agcaccacga ggcactggtc ggccacgggt ttacacacgc gcacatcgtt
     1141 gcgttaagcc aacacccggc agcgttaggg accgtcgctg tcaagtatca ggacatgatc
     1201 gcagcgttgc cagaggcgac acacgaagcg atcgttggcg tcggcaaaca gtggtccggc
     1261 gcacgcgctc tggaggcctt gctcacggtg gcgggagagt tgagaggtcc accgttacag
     1321 ttggacacag gccaacttct caagattgca aaacgtggcg gcgtgaccgc agtggaggca
     1381 gtgcatgcat ggcgcaatgc actgacgggt gccccgctca aggagacggg cgccgctaca
     1441 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     1501 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     1561 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     1621 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcctcc agcttttgtt
     1681 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1741 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1801 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1861 tccaccggtc gtctccaccc ctgagcaggt agtggctatt gcatccaaca acgggggcag
     1921 acccgcactg gagtcaattg ttgcccagtt atctcgccct gatccggcgt tggccgcgtt
     1981 gaccaacgac cacctcgtcg ccttggcctg cctcggcggg cgtcctgcgc tggatgcagt
     2041 gaaaaaggga ttggggggtt ctgcgattcc agttaagaga ggagctacag gagaaacgaa
     2101 agtctttact gggaattcca attctcccaa atcaccgact aaaggcggat gtagtggtgg
     2161 tagtaccaat ctttccgaca ttatcgagaa ggaaacaggt aaacaactcg taatccaaga
     2221 aagcatactg atgcttcctg aagaggttga agaggtcata gggaacaaac ctgaaagcga
     2281 cattttggtt catactgcct atgatgagtc tacagatgag aacgtgatgt tgctaacctc
     2341 agatgcacct gaatacaagc catgggcttt agtgattcag gattcgaatg gagagaacaa
     2401 gatcaagatg ctctgataac tcgagcgatc ctctagaacc cagctttctt gtacaaagtt
     2461 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2521 aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct atagtgagtc
     2581 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2641 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2701 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2761 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2821 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2881 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2941 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3001 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3061 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3121 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctt gctcaggcgc aatcacgaat
     3181 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3241 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3301 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3361 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3421 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3481 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3541 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3601 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4021 cttggagcga acg
//
