LOCUS       sequence-413166-        3363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Guide RNA expression cassette harboring AtU3 promoter, ge4.1 5' 
            modification, and ge4.1 3' modification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3363)
  AUTHORS   Tang X, Eid A, Zhang R, Cheng Y, Liu A, Chen Y, Chen P, Zhang Y, Qi 
            Y
  TITLE     Genome editing in rice and tomato with a small Un1Cas12f1 nuclease.
  JOURNAL   Plant Genome. 2024 May 28:e20465. doi: 10.1002/tpg2.20465.
  PUBMED    38807445
REFERENCE   2  (bases 1 to 3363)
  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 
            Genome. 2024 May 28:e20465. doi: 10.1002/tpg2.20465."
COMMENT     Sequence Label: pYPQ141B-ge4.1
FEATURES             Location/Qualifiers
     source          1..3363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(536..635)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(653..671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(688..706)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(711..727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(711..727)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1158..1949
                     /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      2042..2630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2531..2550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2784..2801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2960..2987)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3079..3165)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3214..3236
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3228..3245
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3229..3245
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3260..3355
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cgttccctct agataacgca ggatcctatt tactttaaat ttttcttatg gctcagcctg
       61 tgatggataa ctgaatcaaa caaatggcgt ctgggtttaa gaacatctgt tttggctatg
      121 ttggacgaaa caagtgaact tttaggatca acttccgttt atatacggag cttatatcga
      181 gcaataagat aagtgggctt tttatgtaat ttaatgggct atcgtccata tattcactaa
      241 tacccatgcc cagtacccat gtatgcgttt catataagct cctaatttct cccacatcgc
      301 tcaaatctaa acaaatcttg ttgtatatat aacactgagg gagcaccatt ggtcaccgct
      361 tcacttagag ntgaaggtgg gctgcttgca tcagcctaat gtcgagaagt gctttcttcg
      421 gaaagtaacc ctcgaaacaa agaaaggaat gcaacggaga cgagatctag tctgagtcga
      481 ccgtctcttt ttattttttt tccctttcct tttttaacta gtaagggcga attcgaccca
      541 gctttcttgt acaaagttgg cattataaaa aataattgct catcaatttg ttgcaacgaa
      601 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
      661 gagtcgtatt acatggtcat agcatcccct atagtgagtc gtattacatg gtcatagctg
      721 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
      781 atatatcatc atgcctcctc tggaccagcc aggacagaaa tgcctcgact tcgctgctac
      841 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
      901 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
      961 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     1021 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     1081 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     1141 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     1201 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     1261 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     1321 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     1381 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     1441 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     1501 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     1561 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     1621 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     1681 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     1741 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     1801 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     1861 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     1921 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     1981 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2161 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2701 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2761 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2821 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2881 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gcgagccagg
     2941 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     3001 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     3061 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     3121 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     3181 ctactctcgc gttaacgctt gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
     3241 ccagtcttaa gctcgggccc aataatgatt ttattttgac tgatagtgac ctgttcgttg
     3301 caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa agttgcccca
     3361 tgg
//
