LOCUS       sequence-325389-        8367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone (attL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8367)
  AUTHORS   Ren Q, Sretenovic S, Liu G, Zhong Z, Wang J, Huang L, Tang X, Guo Y,
            Liu L, Wu Y, Zhou J, Zhao Y, Yang H, He Y, Liu S, Yin D, Mayorga R, 
            Zheng X, Zhang T, Qi Y, Zhang Y
  TITLE     Improved plant cytosine base editors with high editing activity, 
            purity, and specificity.
  JOURNAL   Plant Biotechnol J. 2021 May 27. doi: 10.1111/pbi.13635.
  PUBMED    34042262
REFERENCE   2  (bases 1 to 8367)
  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"; journalName: "Plant 
            Biotechnol J. 2021 May 27. doi: 10.1111/pbi.13635."
COMMENT     Sequence Label: pYPQ265F2
FEATURES             Location/Qualifiers
     source          1..8367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             358..1149
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1242..1830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1731..1750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1984..2001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2160..2187)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2279..2365)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2414..2436
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2428..2445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2429..2445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2461..2560
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3227..3292
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3299..3319
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             7357..7365
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7445..7492
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7514..7762
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7775..7795
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7817..8069
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8114..8237
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(8254..8273)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8255..8273)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8278..8294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8278..8294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
       61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
      121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
      181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
      241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
      301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
      361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
      421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
      601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
      721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
      901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
      961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa
     2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt
     2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattcgc ccttcatgga
     2581 ggcctctccg gcttcaggtc ctcgccatct catggacccc catatcttta cctccaattt
     2641 caataacggc attggcaggc acaagacata cctctgttat gaggtcgagc ggctcgataa
     2701 cggaacctca gtgaagatgg accaacacag gggtttcctt cataaccaag cgaaaaatct
     2761 cctttgcggt ttctatgggc ggcacgcgga gctgagattc ttggatttgg tcccgagcct
     2821 ccagctggat cccgcgcaaa tataccgcgt cacttggttc atatcatgga gcccgtgctt
     2881 tagctggggg tgtgccggtg aggttcgcgc gttcctccag gaaaacaccc atgttcgcct
     2941 ccggattttc gcagcgagga ttttcgacta tgatccactt tacaaagagg ccctccaaat
     3001 gcttagagat gctggggctc aagtttcaat tatgacatac gacgagttta agcattgttg
     3061 ggacactttt gtcgatcatc agggttgtcc cttccagccg tgggatggtc ttgatgagca
     3121 ttcgcaagct ttgagtggca ggcttcgcgc gatacttcag aaccagggta actcaggttc
     3181 tgagacccct ggcacaagtg agtcagcaac acccgagtcc accatggatt acaaggacca
     3241 cgacggggat tacaaggacc acgacattga ttacaaggat gatgatgaca agatggctcc
     3301 gaagaagaag aggaaggttg gcatccacgg ggtgccagct gctgacaaga agtactcgat
     3361 cggcctcgct attgggacta actctgttgg ctgggccgtg atcaccgacg agtacaaggt
     3421 gccctcaaag aagttcaagg tcctgggcaa caccgatcgg cattccatca agaagaatct
     3481 cattggcgct ctcctgttcg acagcggcga gacggctgag gctacgcggc tcaagcgcac
     3541 cgcccgcagg cggtacacgc gcaggaagaa tcgcatctgc tacctgcagg agattttctc
     3601 caacgagatg gcgaaggttg acgattcttt cttccacagg ctggaggagt cattcctcgt
     3661 ggaggaggat aagaagcacg agcggcatcc aatcttcggc aacattgtcg acgaggttgc
     3721 ctaccacgag aagtacccta cgatctacca tctgcggaag aagctcgtgg actccacaga
     3781 taaggcggac ctccgcctga tctacctcgc tctggcccac atgattaagt tcaggggcca
     3841 tttcctgatc gagggggatc tcaacccgga caatagcgat gttgacaagc tgttcatcca
     3901 gctcgtgcag acgtacaacc agctcttcga ggagaacccc attaatgcgt caggcgtcga
     3961 cgcgaaggct atcctgtccg ctaggctctc gaagtctcgg cgcctcgaga acctgatcgc
     4021 ccagctgccg ggcgagaaga agaacggcct gttcgggaat ctcattgcgc tcagcctggg
     4081 gctcacgccc aacttcaagt cgaatttcga tctcgctgag gacgccaagc tgcagctctc
     4141 caaggacaca tacgacgatg acctggataa cctcctggcc cagatcggcg atcagtacgc
     4201 ggacctgttc ctcgctgcca agaatctgtc ggacgccatc ctcctgtctg atattctcag
     4261 ggtgaacacc gagattacga aggctccgct ctcagcctcc atgatcaagc gctacgacga
     4321 gcaccatcag gatctgaccc tcctgaaggc gctggtcagg cagcagctcc ccgagaagta
     4381 caaggagatc ttcttcgatc agtcgaagaa cggctacgct gggtacattg acggcggggc
     4441 ctctcaggag gagttctaca agttcatcaa gccgattctg gagaagatgg acggcacgga
     4501 ggagctgctg gtgaagctca atcgcgagga cctcctgagg aagcagcgga cattcgataa
     4561 cggcagcatc ccacaccaga ttcatctcgg ggagctgcac gctatcctga ggaggcagga
     4621 ggacttctac cctttcctca aggataaccg cgagaagatc gagaagattc tgactttcag
     4681 gatcccgtac tacgtcggcc cactcgctag gggcaactcc cgcttcgctt ggatgacccg
     4741 caagtcagag gagacgatca cgccgtggaa cttcgaggag gtggtcgaca agggcgctag
     4801 cgctcagtcg ttcatcgaga ggatgacgaa tttcgacaag aacctgccaa atgagaaggt
     4861 gctccctaag cactcgctcc tgtacgagta cttcacagtc tacaacgagc tgactaaggt
     4921 gaagtatgtg accgagggca tgaggaagcc ggctttcctg tctggggagc agaagaaggc
     4981 catcgtggac ctcctgttca agaccaaccg gaaggtcacg gttaagcagc tcaaggagga
     5041 ctacttcaag aagattgagt gcttcgattc ggtcgagatc tctggcgttg aggaccgctt
     5101 caacgcctcc ctggggacct accacgatct cctgaagatc attaaggata aggacttcct
     5161 ggacaacgag gagaatgagg atatcctcga ggacattgtg ctgacactca ctctgttcga
     5221 ggaccgggag atgatcgagg agcgcctgaa gacttacgcc catctcttcg atgacaaggt
     5281 catgaagcag ctcaagagga ggaggtacac cggctggggg aggctgagca ggaagctcat
     5341 caacggcatt cgggacaagc agtccgggaa gacgatcctc gacttcctga agagcgatgg
     5401 cttcgcgaac cgcaatttca tgcagctgat tcacgatgac agcctcacat tcaaggagga
     5461 tatccagaag gctcaggtga gcggccaggg ggactcgctg cacgagcata tcgcgaacct
     5521 cgctggctcg ccagctatca agaaggggat tctgcagacc gtgaaggttg tggacgagct
     5581 ggtgaaggtc atgggcaggc acaagcctga gaacatcgtc attgagatgg cccgggagaa
     5641 tcagaccacg cagaagggcc agaagaactc acgcgagagg atgaagagga tcgaggaggg
     5701 cattaaggag ctggggtccc agatcctcaa ggagcacccg gtggagaaca cgcagctgca
     5761 gaatgagaag ctctacctgt actacctcca gaatggccgc gatatgtatg tggaccagga
     5821 gctggatatt aacaggctca gcgattacga cgtcgatcat atcgttccac agtcattcct
     5881 gaaggatgac tccattgaca acaaggtcct caccaggtcg gacaagaacc ggggcaagtc
     5941 tgataatgtt ccttcagagg aggtcgttaa gaagatgaag aactactggc gccagctcct
     6001 gaatgccaag ctgatcacgc agcggaagtt cgataacctc acaaaggctg agaggggcgg
     6061 gctctctgag ctggacaagg cgggcttcat caagaggcag ctggtcgaga cacggcagat
     6121 cactaagcac gttgcgcaga ttctcgactc acggatgaac actaagtacg atgagaatga
     6181 caagctgatc cgcgaggtga aggtcatcac cctgaagtca aagctcgtct ccgacttcag
     6241 gaaggatttc cagttctaca aggttcggga gatcaacaat taccaccatg cccatgacgc
     6301 gtacctgaac gcggtggtcg gcacagctct gatcaagaag tacccaaagc tcgagagcga
     6361 gttcgtgtac ggggactaca aggtttacga tgtgaggaag atgatcgcca agtcggagca
     6421 ggagattggc aaggctaccg ccaagtactt cttctactct aacattatga atttcttcaa
     6481 gacagagatc actctggcca atggcgagat ccggaagcgc cccctcatcg agacgaacgg
     6541 cgagacgggg gagatcgtgt gggacaaggg cagggatttc gcgaccgtca ggaaggttct
     6601 ctccatgcca caagtgaata tcgtcaagaa gacagaggtc cagactggcg ggttctctaa
     6661 ggagtcaatt cgccctaagc ggaacagcga caagctcatc gcccgcaaga aggactggga
     6721 tccgaagaag tacggcgggt tcgttagccc cactgtggcc tactcggtcc tggttgtggc
     6781 gaaggttgag aagggcaagt ccaagaagct caagagcgtg aaggagctgc tggggatcac
     6841 gattatggag cgctccagct tcgagaagaa cccgatcgat ttcctggagg cgaagggcta
     6901 caaggaggtg aagaaggacc tgatcattaa gctccccaag tactcactct tcgagctgga
     6961 gaacggcagg aagcggatgc tggcttccgc tcgcttcctg cagaagggga acgagctggc
     7021 tctgccgtcc aagtatgtga acttcctcta cctggcctcc cactacgaga agctcaaggg
     7081 cagccccgag gacaacgagc agaagcagct gttcgtcgag cagcacaagc attacctcga
     7141 cgagatcatt gagcagattt ccgagttctc caagcgcgtg atcctggccg acgcgaatct
     7201 ggataaggtc ctctccgcgt acaacaagca ccgcgacaag ccaatcaggg agcaggctga
     7261 gaatatcatt catctcttca ccctgacgaa cctcggcgcc cctcgcgctt tcaagtactt
     7321 cgacacaact atcgatcgca aggtctaccg cagcactaag gaggtcctgg acgcgaccct
     7381 catccaccag tcgattaccg gcctctacga gacgcgcatc gacctgtctc agctcggggg
     7441 cgacaagcgg ccagcggcga cgaagaaggc ggggcaggcg aagaagaaga agggagacgg
     7501 atcaggcggg tcaacaaacc tcagtgacat catagagaag gaaactggta agcaactggt
     7561 tattcaagag agcattctga tgctccctga ggaagtcgag gaagttatag gaaacaagcc
     7621 tgagagcgat atactcgtgc atacggcgta tgacgagagc acggatgaaa atgtgatgct
     7681 cttgaccagc gatgcgccag aatataaacc ctgggcactg gttattcagg actctaacgg
     7741 ggaaaataaa atcaagatgc tctcaggtgg ctccccaaag aagaaacgca aggtttagtg
     7801 gcggccgccg acgtccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     7861 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     7921 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     7981 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     8041 gcgcgcggtg tcatctatgt tactagatcg ggaattgatc ccccctcgac agcttccgga
     8101 aagggcgaat tcgcaacttt gtatacaaaa gttgaacgag aaacgtaaaa tgatataaat
     8161 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     8221 catatccagt cactatgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     8281 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     8341 gataaaaata tatcatcatg cctcctc
//
