LOCUS       sequence-343426-        8871 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 8871)
  AUTHORS   Sretenovic S, Liu S, Li G, Cheng Y, Fan T, Xu Y, Zhou J, Zheng X, 
            Coleman G, Zhang Y, Qi Y
  TITLE     Exploring C-To-G Base Editing in Rice, Tomato, and Poplar.
  JOURNAL   Front Genome Ed. 2021 Sep 15;3:756766. doi: 
            10.3389/fgeed.2021.756766. eCollection 2021.
  PUBMED    34713268
REFERENCE   2  (bases 1 to 8871)
  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: "Front 
            Genome Ed."; date: "2021-09-15"; volume: "3:756766. doi"; pages: " 
            10.3389/fgeed.2021.756766. eCollection 2021"
COMMENT     Sequence Label: pYPQ265N2
FEATURES             Location/Qualifiers
     source          1..8871
                     /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             2576..3262
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             3317..3382
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3389..3409
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             7447..7455
                     /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             7535..7582
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8279..8299
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8321..8573
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8618..8741
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(8758..8777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8759..8777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8782..8798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8782..8798)
                     /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 ccttcatgag
     2581 ttctgaaaca ggtccagtgg ctgtcgatcc gactctgagg aggcgcatcg aaccacacga
     2641 atttgaggtt ttttttgatc caagagagct tcgcaaggaa acttgtctcc tgtatgagat
     2701 aaattgggga ggaagacata gtatctggag acacacaagc cagaacacca ataagcatgt
     2761 cgaagttaac ttcatcgaaa aatttacgac cgaacgctac ttttgtccta atacacgctg
     2821 cagtatcact tggttccttt cgtggtcacc ttgcggtgaa tgttccagag caattacgga
     2881 gtttctctcg agatatccac acgttacact tttcatttac atagctcggc tttatcatca
     2941 cgcagatccg agaaatcgcc aggggctgcg cgatctgatt tcgtccggtg tcaccattca
     3001 aattatgact gagcaggaat ctggatactg ctggcgcaac ttcgtgaact acagtccgtc
     3061 gaatgaagcg cactggccga gatatcccca tctctgggtg cgcttgtatg ttttggagct
     3121 ttattgtatc atcttgggat tgccaccttg tcttaatata ttgcgccgga aacaaccgca
     3181 gctcactttt ttcaccattg ctctgcagtc ttgtcactac caacgcctcc ccccgcacat
     3241 cctgtgggcc acaggcctta aatcaggttc tgagacccct ggcacaagtg agtcagcaac
     3301 acccgagtcc accatggatt acaaggacca cgacggggat tacaaggacc acgacattga
     3361 ttacaaggat gatgatgaca agatggctcc gaagaagaag aggaaggttg gcatccacgg
     3421 ggtgccagct gctgacaaga agtactcgat cggcctcgct attgggacta actctgttgg
     3481 ctgggccgtg atcaccgacg agtacaaggt gccctcaaag aagttcaagg tcctgggcaa
     3541 caccgatcgg cattccatca agaagaatct cattggcgct ctcctgttcg acagcggcga
     3601 gacggctgag aggacgcggc tcaagcgcac cgcccgcagg cggtacacgc gcaggaagaa
     3661 tcgcatctgc tacctgcagg agattttctc caacgagatg gcgaaggttg acgattcttt
     3721 cttccacagg ctggaggagt cattcctcgt ggaggaggat aagaagcacg agcggcatcc
     3781 aatcttcggc aacattgtcg acgaggttgc ctaccacgag aagtacccta cgatctacca
     3841 tctgcggaag aagctcgtgg actccacaga taaggcggac ctccgcctga tctacctcgc
     3901 tctggcccac atgattaagt tcaggggcca tttcctgatc gagggggatc tcaacccgga
     3961 caatagcgat gttgacaagc tgttcatcca gctcgtgcag acgtacaacc agctcttcga
     4021 ggagaacccc attaatgcgt caggcgtcga cgcgaaggct atcctgtccg ctaggctctc
     4081 gaagtctcgg cgcctcgaga acctgatcgc ccagctgccg ggcgagaaga agaacggcct
     4141 gttcgggaat ctcattgcgc tcagcctggg gctcacgccc aacttcaagt cgaatttcga
     4201 tctcgctgag gacgccaagc tgcagctctc caaggacaca tacgacgatg acctggataa
     4261 cctcctggcc cagatcggcg atcagtacgc ggacctgttc ctcgctgcca agaatctgtc
     4321 ggacgccatc ctcctgtctg atattctcag ggtgaacacc gagattacga aggctccgct
     4381 ctcagcctcc atgatcaagc gctacgacga gcaccatcag gatctgaccc tcctgaaggc
     4441 gctggtcagg cagcagctcc ccgagaagta caaggagatc ttcttcgatc agtcgaagaa
     4501 cggctacgct gggtacattg acggcggggc ctctcaggag gagttctaca agttcatcaa
     4561 gccgattctg gagaagatgg acggcacgga ggagctgctg gtgaagctca atcgcgagga
     4621 cctcctgagg aagcagcgga cattcgataa cggcagcatc ccacaccaga ttcatctcgg
     4681 ggagctgcac gctatcctga ggaggcagga ggacttctac cctttcctca aggataaccg
     4741 cgagaagatc gagaagattc tgactttcag gatcccgtac tacgtcggcc cactcgctag
     4801 gggcaactcc cgcttcgctt ggatgacccg caagtcagag gagacgatca cgccgtggaa
     4861 cttcgaggag gtggtcgaca agggcgctag cgctcagtcg ttcatcgaga ggatgacgaa
     4921 tttcgacaag aacctgccaa atgagaaggt gctccctaag cactcgctcc tgtacgagta
     4981 cttcacagtc tacaacgagc tgactaaggt gaagtatgtg accgagggca tgaggaagcc
     5041 ggctttcctg tctggggagc agaagaaggc catcgtggac ctcctgttca agaccaaccg
     5101 gaaggtcacg gttaagcagc tcaaggagga ctacttcaag aagattgagt gcttcgattc
     5161 ggtcgagatc tctggcgttg aggaccgctt caacgcctcc ctggggacct accacgatct
     5221 cctgaagatc attaaggata aggacttcct ggacaacgag gagaatgagg atatcctcga
     5281 ggacattgtg ctgacactca ctctgttcga ggaccgggag atgatcgagg agcgcctgaa
     5341 gacttacgcc catctcttcg atgacaaggt catgaagcag ctcaagagga ggaggtacac
     5401 cggctggggg aggctgagca ggaagctcat caacggcatt cgggacaagc agtccgggaa
     5461 gacgatcctc gacttcctga agagcgatgg cttcgcgaac cgcaatttca tgcagctgat
     5521 tcacgatgac agcctcacat tcaaggagga tatccagaag gctcaggtga gcggccaggg
     5581 ggactcgctg cacgagcata tcgcgaacct cgctggctcg ccagctatca agaaggggat
     5641 tctgcagacc gtgaaggttg tggacgagct ggtgaaggtc atgggcaggc acaagcctga
     5701 gaacatcgtc attgagatgg cccgggagaa tcagaccacg cagaagggcc agaagaactc
     5761 acgcgagagg atgaagagga tcgaggaggg cattaaggag ctggggtccc agatcctcaa
     5821 ggagcacccg gtggagaaca cgcagctgca gaatgagaag ctctacctgt actacctcca
     5881 gaatggccgc gatatgtatg tggaccagga gctggatatt aacaggctca gcgattacga
     5941 cgtcgatcat atcgttccac agtcattcct gaaggatgac tccattgaca acaaggtcct
     6001 caccaggtcg gacaagaacc ggggcaagtc tgataatgtt ccttcagagg aggtcgttaa
     6061 gaagatgaag aactactggc gccagctcct gaatgccaag ctgatcacgc agcggaagtt
     6121 cgataacctc acaaaggctg agaggggcgg gctctctgag ctggacaagg cgggcttcat
     6181 caagaggcag ctggtcgaga cacggcagat cactaagcac gttgcgcaga ttctcgactc
     6241 acggatgaac actaagtacg atgagaatga caagctgatc cgcgaggtga aggtcatcac
     6301 cctgaagtca aagctcgtct ccgacttcag gaaggatttc cagttctaca aggttcggga
     6361 gatcaacaat taccaccatg cccatgacgc gtacctgaac gcggtggtcg gcacagctct
     6421 gatcaagaag tacccaaagc tcgagagcga gttcgtgtac ggggactaca aggtttacga
     6481 tgtgaggaag atgatcgcca agtcggagca ggagattggc aaggctaccg ccaagtactt
     6541 cttctactct aacattatga atttcttcaa gacagagatc actctggcca atggcgagat
     6601 ccggaagcgc cccctcatcg agacgaacgg cgagacgggg gagatcgtgt gggacaaggg
     6661 cagggatttc gcgaccgtca ggaaggttct ctccatgcca caagtgaata tcgtcaagaa
     6721 gacagaggtc cagactggcg ggttctctaa ggagtcaatt aggcctaagc ggaacagcga
     6781 caagctcatc gcccgcaaga aggactggga tccgaagaag tacggcgggt tcctgtggcc
     6841 cactgtggcc tactcggtcc tggttgtggc gaaggttgag aagggcaagt ccaagaagct
     6901 caagagcgtg aaggagctgc tggggatcac gattatggag cgctccagct tcgagaagaa
     6961 cccgatcgat ttcctggagg cgaagggcta caaggaggtg aagaaggacc tgatcattaa
     7021 gctccccaag tactcactct tcgagctgga gaacggcagg aagcggatgc tggcttccgc
     7081 taagcagctg cagaagggga acgagctggc tctgccgtcc aagtatgtga acttcctcta
     7141 cctggcctcc cactacgaga agctcaaggg cagccccgag gacaacgagc agaagcagct
     7201 gttcgtcgag cagcacaagc attacctcga cgagatcatt gagcagattt ccgagttctc
     7261 caagcgcgtg atcctggccg acgcgaatct ggataaggtc ctctccgcgt acaacaagca
     7321 ccgcgacaag ccaatcaggg agcaggctga gaatatcatt catctcttca ccctgacgag
     7381 gctcggcgcc cctagggctt tcaagtactt cgacacaact atcgatccga agcagtacag
     7441 gagcactaag gaggtcctgg acgcgaccct catccaccag tcgattaccg gcctctacga
     7501 gacgcgcatc gacctgtctc agctcggggg cgacaagcgg ccagcggcga cgaagaaggc
     7561 ggggcaggcg aagaagaaga agggagacgg atcaggcggg tcaatgtttg gagagagctg
     7621 gaagaagcac ctcagcgggg agttcgggaa accgtatttt atcaagctaa tgggatttgt
     7681 tgcagaagaa agaaagcatt acactgttta tccaccccca caccaagtct tcacctggac
     7741 ccagatgtgt gacataaaag atgtgaaggt tgtcatcctg ggacaggatc catatcatgg
     7801 acctaatcaa gctcacgggc tctgctttag tgttcaaaga cctgttccgc ctccgcccag
     7861 tttggagaac atttacaaag agttgtctac agacatagag gattttgttc atcctggcca
     7921 cggagattta tctgggtggg ccaagcaagg tgttctcctt ctcaacgctg tcctcacggt
     7981 tcgtgcccat caagccaact ctcataagga gcgaggctgg gagcagttca ctgatgcagt
     8041 tgtgtcctgg ctaaatcaga actcgaatgg ccttgttttc ttgctctggg gctcttatgc
     8101 tcagaagaag ggcagtgcca ttgataggaa gcggcaccat gtactacaga cggctcatcc
     8161 ctcccctttg tcagtgtata gagggttctt tggatgtaga cacttttcaa agaccaatga
     8221 gctgctgcag aagtctggca agaagcccat tgactggaag gagctgtcag gtggctcccc
     8281 aaagaagaaa cgcaaggttt agtggcggcc gccgacgtcc gatcgttcaa acatttggca
     8341 ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct
     8401 gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg
     8461 ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata
     8521 gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt
     8581 gatcccccct cgacagcttc cggaaagggc gaattcgcaa ctttgtatac aaaagttgaa
     8641 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
     8701 actacataat actgtaaaac acaacatatc cagtcactat gccatccagc tgatatcccc
     8761 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     8821 aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct c
//
