LOCUS       sequence-338715-        6730 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 6730)
  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 6730)
  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_NI_G1397-DddtoxA-C
FEATURES             Location/Qualifiers
     source          1..6730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     953..972
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(992..1069)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1115..1134
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1115..1133
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1134..1158
                     /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)."
     CDS             1211..1240
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1250..1267
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1280..1573
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             4398..4442
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4480..4498)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4494..4541
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4771..5586)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4771..5586)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5494..5513
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5587..5678)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5694..6443)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(5704..5721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5944..5963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6639..6730,1..991))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
       61 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
      121 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
      181 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
      241 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
      301 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
      361 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
      421 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
      481 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
      541 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
      601 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
      661 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
      721 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
      781 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
      841 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
      901 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
      961 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     1021 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     1081 cgctctccct tatgcgactc ctgcattagg aaattaatac gactcactat aggggaattg
     1141 tgagcggata acaattcccc tgtagaaata attttgttta actttaataa ggagatatac
     1201 catgggcagc caccaccacc accatcatca tcatcatcac agcagcggcc tggtgccgcg
     1261 cggcagccac atggcaagca tgtcggactc agaagtcaat caagaagcta agccagaggt
     1321 caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga
     1381 gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa
     1441 aagacagggt aaggaaatgg actccttaag attcttgtac gacggtatta gaatccaagc
     1501 tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga
     1561 acagattggt ggatccgccg acgcgcagtc ttttctgaac cgcgtatgcg gggtaagcgc
     1621 agctcgcttg acaccctgtg gtactgggac gtctacagac gtggtttatc gcgcgttcga
     1681 catctataac gacaaagtcg ccggtttcgc taagttcttg aagaccaatt gctgtcgctt
     1741 tcaagagaaa gatgaagatg ataatcttat tgactcttat tttgtagtaa agcgtcacac
     1801 cttcagcaac tatcagcatg aagaaaccat ctataatctt ttgaaggact gtccagcagt
     1861 agccaagcac gattttttca aatttcgtat tgacggtgac atggtccccc acatttcacg
     1921 ccagcgcctt accaaataca caatggcgga tctggtttat gctcttcgtc attttgacga
     1981 aggaaattgc gacaccctta aggagattct ggtaacttat aattgttgcg acgatgacta
     2041 ttttaataaa aaggattggt atgattttgt cgaaaatccg gatattttac gtgtgtacgc
     2101 aaatctggga gagcgcgttc gccaagcgtt gttaaagacg gtacagttct gcgacgccat
     2161 gcgtaatgcc ggtatcgtgg gtgtgctgac cttagataac caagatctta acggtaattg
     2221 gtatgatttc ggagatttta ttcagaccac tcccggctcc ggcgtgccgg tagtggattc
     2281 ctattatagt ctgctgatgc caatcttgac gttgacccgt gccttgacgg cggagagcca
     2341 tgtggacaca gacttaacga agccttacat taaatgggat ttgcttaaat acgacttcac
     2401 ggaggagcgt cttaaacttt ttgaccgtta ctttaagtat tgggaccaga cgtatcaccc
     2461 caactgcgtc aactgcttag acgatcgctg catcttacat tgtgcaaact ttaatgtttt
     2521 attttccaca gtttttccgc cgacttcctt cgggccactg gtacgcaaga tttttgtcga
     2581 cggcgtgccg tttgtagtct ctacggggta ccactttcgt gaacttggtg ttgttcataa
     2641 ccaggatgtt aatttacatt cgtcacgtct ttcattcaaa gagttactgg tatacgcggc
     2701 cgatcccgcc atgcacgcag cgtccgggaa tttacttctg gataaacgta ctacgtgttt
     2761 cagtgttgca gcgctgacca acaatgtagc tttccaaaca gtaaaaccag ggaatttcaa
     2821 taaggatttt tacgacttcg cagtcagtaa gggttttttt aaagagggga gctcggttga
     2881 gttaaaacac ttttttttcg ctcaggatgg gaacgctgca atctccgatt acgactacta
     2941 tcgctacaac ttacctacga tgtgcgatat tcgccaatta ctgttcgtag ttgaagtagt
     3001 agacaagtat ttcgactgtt acgatggagg atgtattaat gcgaaccaag taatcgtcaa
     3061 taatttagat aaatctgcag gttttccctt taataagtgg ggaaaggcgc gcctgtacta
     3121 cgattcaatg tcgtatgagg accaggacgc tcttttcgca tataccaaac gcaatgttat
     3181 cccgacgatt acgcaaatga atctgaaata tgccatttcc gctaaaaatc gcgctcgcac
     3241 ggtcgcggga gtatcgattt gttctactat gactaaccgc caattccatc aaaagttgct
     3301 taaatctatc gcagcaacgc gtggggcaac ggtggtcatc gggacctcga agttttatgg
     3361 aggttggcac aacatgctga aaaccgtgta tagtgacgtg gaaaacccgc atctgatggg
     3421 gtgggattat ccgaaatgcg atcgcgcaat gcctaatatg cttcgcatca tggcctcttt
     3481 agtgctagca cgtaagcata cgacatgttg cagcttgtcc catcgctttt accgtttagc
     3541 taatgagtgc gcacaagtat tatcggagat ggtaatgtgc ggcgggagtt tatacgtgaa
     3601 gccaggtggt actagttctg gggatgcaac cacagcgtat gctaattcgg ttttcaatat
     3661 ctgtcaggcc gttaccgcca acgtaaacgc acttctgtca acggatggta ataaaattgc
     3721 tgataagtac gtacgcaacc tgcaacatcg tctgtatgag tgcctttatc gcaaccgcga
     3781 tgtcgacacc gactttgtaa acgagttcta tgcatatttg cgtaagcatt tctctatgat
     3841 gatcctttca gacgatgccg ttgtgtgttt caacagtacc tatgcctcgc aaggcctagt
     3901 agcctccatc aaaaacttta agtccgttct ttactatcag aacaacgtct ttatgtcaga
     3961 ggccaagtgc tggaccgaga cggacttaac taaaggccca cacgagttct gctcacaaca
     4021 tactatgctt gtaaagcaag gtgatgacta cgtctatctt ccttatcctg acccaagccg
     4081 catcctggga gctggttgtt tcgttgacga tatcgtgaag accgatggca ccctgatgat
     4141 tgagcgtttc gtctctctgg cgattgacgc gtatccgtta actaagcacc ccaatcaaga
     4201 atacgcggac gtttttcact tatacttaca gtatatccgt aagttacacg atgagctgac
     4261 cggtcacatg cttgatatgt acagcgttat gctgacgaat gacaacactt cgcgttattg
     4321 ggagcctgag ttttatgaag cgatgtatac acctcacaca gttctgcagt aagcttgcgg
     4381 ccgcactcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact
     4441 cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat
     4501 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
     4561 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     4621 aagcggctat ttaacgaccc tgccctgaac cgacgacaag ctgacgaccg ggtctccgca
     4681 agtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4741 tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa
     4801 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     4861 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     4921 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     4981 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     5041 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     5101 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct
     5161 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     5221 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     5281 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     5341 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     5401 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     5461 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     5521 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     5581 gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggcatgcagc gctcttccgc
     5701 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg tgtcagctca
     5761 ctcaaaagcg gtaatacggt tatccacaga atcaggggat aaagccggaa agaacatgtg
     5821 agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc gtttttccat aggctccgcc
     5881 cccctgacga gcatcacaaa aatcgacgct caagccagag gtggcgaaac ccgacaggac
     5941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6061 gctcacgctg ttggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6181 acccggtaag acacgactta tcgccactgg cagcagccat tggtaactga tttagaggac
     6241 tttgtcttga agttatgcac ctgttaaggc taaactgaaa gaacagattt tggtgagtgc
     6301 ggtcctccaa cccacttacc ttggttcaaa gagttggtag ctcagcgaac cttgagaaaa
     6361 ccaccgttgg tagcggtggt ttttctttat ttatgagatg atgaatcaat cggtctatca
     6421 agtcaacgaa cagctattcc gttactctag atttcagtgc aatttatctc ttcaaatgta
     6481 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg
     6541 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     6601 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6721 gtattgggcg
//
