LOCUS       sequence-338617-        4244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ptpTALECD vector assembly, Entry clone 1 with attL1 and L4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4244)
  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 4244)
  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: E1_pENTR_L1-L4_NN_G1397-DddtoxA-N
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             523..774
                     /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(800..895)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(921..939)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(944..960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1062..1081)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1073..1882
                     /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(1140..1159)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2029..2617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2518..2537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2771..2788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2947..2974)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3066..3152)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3216..3232
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3248..3347
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3350..3376
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3683..4093)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLLER
                     GTRQCIAPCMHCLHCGHAATFCNLEKLACVQL"
     primer_bind     3958..3974
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3984..4002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4050..4070)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4050..4068)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4089..4105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4102..4124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4113..4129
                     /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(4137..4143,4144..4161,4162..4167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4182..4203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 acccctgagc aggtagtggc tattgcatcc aacaacgggg gcagacccgc actggagtca
       61 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
      121 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
      181 gggtctggat cgtatgcttt aggaccgtat cagatctcag ctccacaatt gcctgcatat
      241 aacggacaaa ctgttgggac cttttactac gttaacgatg ctggtggatt ggagtccaaa
      301 gtgttctctt ctggtggccc aactccatat cccaattatg cgaatgcagg ccatgttgaa
      361 ggtcaatcag ccctattcat gagagataac ggaataagtg aaggactggt gtttcacaac
      421 aatccagaag gtacttgtgg attttgcgta aacatgactg agacacttct cccagaaaat
      481 gccaagatga cagttgtacc tcctgaaggt tctggtggat cgacaaacct ttcagacatt
      541 atcgagaaag agacaggcaa acagctagtg attcaagagt ccattctcat gcttcccgaa
      601 gaagttgagg aagtcattgg gaataagccg gaaagtgaca tactcgttca tacggcttac
      661 gatgagagca cggatgagaa tgtcatgttg cttaccagtg atgcacctga atacaaacct
      721 tgggctttag tcatccagga cagcaatggt gagaacaaga tcaagatgct gtgataactc
      781 gagcgatcct ctagacaccc aacttttcta tacaaagttg gcattataag aaagcattgc
      841 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
      901 ccagctgcag ggcggccgcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
      961 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1021 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     1081 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     1141 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     1201 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1261 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1321 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     1381 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1441 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     1501 cgtatttcgc cttgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     1561 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     1621 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     1681 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     1741 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     1801 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     1861 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     1921 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     1981 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2041 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2101 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2161 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2221 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2281 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2341 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2401 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2461 caggtatccg gtaagcggca tggtcggaac aggagagcgc acgagggagc ttccaggggg
     2521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2581 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2641 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2701 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2761 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     2821 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2881 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     2941 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3001 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3061 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3121 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     3181 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     3241 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     3301 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctttt acccatacga
     3361 tgttccagat tacgctgata tcgccgatct acgcacgctc ggctacagcc agcagcaaca
     3421 ggagaagatc aaaccgaagg ttcgttcgac agtggcgcag caccacgagg cactggtcgg
     3481 ccacgggttt acacacgcgc acatcgttgc gttaagccaa cacccggcag cgttagggac
     3541 cgtcgctgtc aagtatcagg acatgatcgc agcgttgcca gaggcgacac acgaagcgat
     3601 cgttggcgtc ggcaaacagt ggtccggcgc acgcgctctg gaggccttgc tcacggtggc
     3661 gggagagttg agaggtccac cgttacagtt ggacacaggc caacttctca agattgcaaa
     3721 acgtggcggc gtgaccgcag tggaggcagt gcatgcatgg cgcaatgcac tgacgggtgc
     3781 cccgctcaag gagacgggcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     3841 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     3901 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     3961 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
     4021 cccccctcga ggtcctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     4081 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4141 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4201 ttaattgcgt tgcgctcact gcccgctttc caccggtcgt ctcc
//
