LOCUS       sequence-474023-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for A3A/Y130F-CBE_V01 based C-to-T base editing for 
            monocot plants; NGG PAM; wide working window; A3A/Y130F-CBE_V01 was 
            driven by ZmUbi1 and the sgRNA was driven by OsU3 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7369)
  AUTHORS   Cheng Y, Li G, Qi A, Mandlik R, Pan C, Wang D, Ge S, Qi Y
  TITLE     A comprehensive all-in-one CRISPR toolbox for large-scale screens in
            plants.
  JOURNAL   Plant Cell. 2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081.
  PUBMED    40202185
REFERENCE   2  (bases 1 to 7369)
  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 Cell.
            2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081."
COMMENT     Sequence Label: pYPQAT16
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1209..1233
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     1421..1443
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1435..1452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1436..1452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2753..3547
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3634..4222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4123..4142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4376..4393
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4408..4548
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4474..4493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4634..4656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      4892..5086
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5152..6225)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(6654..7283)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tcaccttttt agacggcggc gctcggtttc ttcagcggcc aagctggccg gccaggccgc
       61 cagcttggca tcagacaaac cggccaggat ttcatgcagc cgcacggttg agacgtgcgc
      121 gggcggctcg aacacgtacc cggccgcgat catctccgcc tcgatctctt cggtaatgaa
      181 aaacggttcg tcctggccgt cctggtgcgg tttcatgctt gttcctcttg gcgttcattc
      241 tcggcggccg ccagggcgtc ggcctcggtc aatgcgtcct aggcaccgcg ccgcctggcc
      301 tcggtgggcg tcacttcctc gctgcgctca agtgcgcggt acagggtcga gcgatgcacg
      361 ccaagcagtg cagccgcctc tttcacggtg cggccttcct ggtcgatcag ctcgcgggcg
      421 tgcgcgatct gtgccggggt gagggtaggg cgggggccaa acttcacgcc tcgggccttg
      481 gcggcctcgc gcccgctccg ggtgcggtcg atgattaggg aacgctcgaa ctcggcaatg
      541 ccggcgaaca cggtcaacac catgcggccg gccggcgtgg tggtgtcggc ccacggctct
      601 gccaggctac gcaggcccgc gccggcctcc tggatgcgct cggcaatgtc cagtaggtcg
      661 cgggtgctgc gggccaggcg gtctagcctg gtcactgtca caacgtcgcc agggcgtagg
      721 tggtcaagca tcctggccag ctccgggcgg tcgcgcctgg tgccggtgat cttctcggaa
      781 aacagcttgg tgcagccggc cgcgtgcagt tcggcccgtt ggttggtcaa gtcctggtcg
      841 tcggtgctga cgcgggcata gcccagcagg ccagcggcgg cgctcttgtt catggcgtaa
      901 tgtctccggt tctagtcgca agtattctac tttatgcgac taaaacacgc gacaagaaaa
      961 cgccaggaaa agggcagggc ggcagcctgt cgcgtaactt aggacttgtg cgacatgtcg
     1021 ttttcagaag acggctgcac tgaacgtcag aagccgactg cactatagca gcggaggggt
     1081 tggatcaaag tactttgatc ccgaggggaa ccctgtggtt ggcatgcaca tacaaatgga
     1141 cgaacggata aaccttttca cgccctttta aatatccgat tattctaata aacgctcttt
     1201 tctcttaggt ttacccgcca atatatcctg tcaaacactg atagtttaaa ctgaaggcgg
     1261 gaaacgacaa tctgatccaa gctcaagctg ctctagcatt cgccattcag gctgcgcaac
     1321 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     1381 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     1441 acgacggcca gtgccaagct tgcatgcctg cagtgcagcg tgacccggtc gtgcccctct
     1501 ctagggaagg tgcgaacaag tccctgatat gagatcatgt ttgtcatctg gagccataga
     1561 acagggttca tcatgagtca tcaacttacc ttcgccgaca gtgaattcag cagtaagcgc
     1621 cgtcagacca gaaaagagat tttcttgtcc cgcatggagc agattctgcc atggcaaaac
     1681 atggtggaag tcatcgagcc gttttacccc aaggctggta atggccggcg accttatccg
     1741 ctggaaacca tgctacgcat tcactgcatg cagcattggt acaacctgag cgatggcgcg
     1801 atggaagatg ctctgtacga aatcgcctcc atgcgtctgt ttgcccggtt atccctggat
     1861 agcgccttgc cggaccgcac caccatcatg aatttccgcc acctgctgga gcagcatcaa
     1921 ctggcccgcc aattgttcaa gaccatcaat cgctggctgg ccgaagcagg cgtcatgatg
     1981 actcaaggca ccttggtcga tgccaccatc attgaggcac ccagctcgac caagaacaaa
     2041 gagcagcaac gcgatccgga gatgcatcag accaagaaag gcaatcagtg gcactttggc
     2101 atgaaggccc acattggtgt cgatgccaag agtggcctga cccacagcct ggtcaccacc
     2161 gcggccaacg agcatgacct caatcagctg ggtaatctgc tgcatggaga ggagcaattt
     2221 gtctcagccg atgccggcta ccaaggggcg ccacagcgcg aggagctggc cgaggtggat
     2281 gtggactggc tgatcgccga gcgccccggc aaggtaagaa ccttgaaaca gcatccacgc
     2341 aagaacaaaa cggccatcaa catcgaatac atgaaagcca gcatccgggc cagggtggag
     2401 cacccatttc gcatcatcaa gcgacagttc ggcttcgtga aagccagata caaggggttg
     2461 ctgaaaaacg ataaccaact ggcgatgtta ttcacgctgg ccaacctgtt tcgggcggac
     2521 caaatgatac gtcagtggga gagatctcac taaaaactgg ggataacgcc ttaaatggcg
     2581 aagaaacggt ctaaataggc tgattcaagg catttacggg agaaaaaatc ggctcaaaca
     2641 tgaagaaatg aaatgactga gtcagccgag aagaatttcc ccgcttattc gcaccttcct
     2701 tagcttcttg gggtatcttt aaatactgta gaaaagagga aggaaataat aaatggctaa
     2761 aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac
     2821 ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt
     2881 aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat
     2941 gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga
     3001 tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga
     3061 agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca
     3121 ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt
     3181 ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg aagaagacac
     3241 tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga
     3301 acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt
     3361 aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt
     3421 ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga
     3481 cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt
     3541 gttttagtac ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3781 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4201 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4261 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4321 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4381 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4441 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4501 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4561 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4621 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4681 gcgaggcagg gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc
     4741 gcgccctggt agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat
     4801 aggccgacgc gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg
     4861 cagctcttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt
     4921 tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt
     4981 cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg
     5041 gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa acagaccttt
     5101 tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc
     5161 ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg
     5221 ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac
     5281 ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa
     5341 caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc
     5401 gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc
     5461 ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc
     5521 ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac
     5581 caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg
     5641 aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt
     5701 gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg
     5761 gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca
     5821 cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta
     5881 gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat
     5941 gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg
     6001 ctcgtcgccc ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga
     6061 ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc
     6121 gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag
     6181 cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc
     6241 ttgggggttc cagtgccatt gcagggccgg caggcaaccc agccgcttac gcctggccaa
     6301 ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt
     6361 tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta
     6421 ctctggtagc tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac
     6481 cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat
     6541 ggctggcgtg tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg
     6601 gccggcactt agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg
     6661 agttttgatt taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt
     6721 tctgattcaa gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg
     6781 atacgggact caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg
     6841 atgcgcgtgc ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg
     6901 acctcaatgc gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg
     6961 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
     7021 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
     7081 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
     7141 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
     7201 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
     7261 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
     7321 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacaca
//
