LOCUS       sequence-492744-        7756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry plasmid (attL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7756)
  AUTHORS   Wang X, Clem J, Malzahn A, Qi Y
  TITLE     Increasing Cas12a- and Cas9-mediated deletion sizes with 
            exonucleases.
  JOURNAL   Plant Physiol. 2025 Oct 31;199(3):kiaf507. doi: 
            10.1093/plphys/kiaf507.
  PUBMED    41182408
REFERENCE   2  (bases 1 to 7756)
  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 
            Physiol."; date: "2025-10-31"; volume: "199(3):kiaf507. doi"; pages:
            " 10.1093/plphys/kiaf507"
COMMENT     Sequence Label: pYPQ230-TREX2-N
FEATURES             Location/Qualifiers
     source          1..7756
                     /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             3294..3341
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3402..3422
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3447..7127
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="SKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAED
                     YKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEI
                     AKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFSE
                     EAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEGE
                     FFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLS
                     DRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNG
                     PAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQL
                     QEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDL
                     LDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSK
                     DKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNGN
                     YEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLI
                     DFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVE
                     EGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKKE
                     ELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKIN
                     TEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDY
                     HSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSGF
                     KNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQN
                     GFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDY
                     KNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQ
                     GDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRNY
                     EAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSV
                     KH"
     CDS             7128..7175
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7206..7458
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7503..7626
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7643..7662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7644..7662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7667..7683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7667..7683)
                     /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 ccttcaccat
     2581 ggcatatgtc agaagcacct cgtgccgaga cctttgtctt cctggacctg gaagccactg
     2641 ggctccccag tgtggagccc gagattgctg agctgtctct atttgctgtc caccgctcct
     2701 ccctggagaa cccggagcac gacgagtctg gtgcactagt attgccacgg gtccttgaca
     2761 agcttacgct gtgcatgtgc cctgagcgcc ccttcactgc taaggctagc gagatcaccg
     2821 gactgagcag tgaaggcctg gcgcgatgtc ggaaggctgg ctttgatggc gcagtggtgc
     2881 ggacactgca ggcattcctg agccgtcagg cagggcccat ctgtcttgtg gcccacaatg
     2941 gatttgatta tgatttccca ctgctgtgtg ccgagctgcg gcgactgggt gctcgactgc
     3001 cccgggatac tgtctgcctg gacactctgc cggcactgcg tggcctggac cgtgcccaca
     3061 gtcacggcac ccgggcacgg ggtcgccagg gttacagtct cggcagtctc ttccatcgct
     3121 acttccgagc agagccaagc gcagcccatt cagctgaggg cgacgtgcat accctgctac
     3181 tgatcttcct gcatcgcgcc gcagagctac tcgcctgggc agatgagcaa gcccgtggat
     3241 gggctcacat cgagcccatg tacttaccgc ctgatgatcc cagcctggag tttaaacgtc
     3301 ctgctgccac caaaaaggcc ggacaggcta agaaaaagaa ggagctctca ggatctgaga
     3361 ccccaggcac aagtgaatca gctacacctg agtccatggc tcctaagaag aagcggaagg
     3421 ttggtattca cggggtgcct gcggcttcaa agctcgagaa attcaccaac tgttattcgt
     3481 tgagcaaaac actgcggttt aaagcgattc cagtcggcaa gactcaagag aatatagaca
     3541 ataagcggct gttggtggaa gatgaaaagc gcgcggaaga ctacaaaggg gtgaagaagt
     3601 tgttggacag atactacctc tcttttatca atgatgtctt gcactcaatc aaattgaaga
     3661 atctgaacaa ctacatctcc ctcttcagaa agaaaacaag gacagaaaag gagaataagg
     3721 aacttgaaaa tttggagatc aatctgagga aagagatcgc gaaagccttt aaaggcaacg
     3781 aaggatacaa aagtctgttc aagaaggata taattgagac aattttgcca gagttcctcg
     3841 atgacaagga cgagattgcg ctggtcaatt cgttcaacgg attcacaaca gcattcacag
     3901 gcttctttga taatcgggaa aatatgttct ctgaggaggc aaagtccact tctattgcgt
     3961 tcaggtgtat caatgagaat ctcactaggt acatttccaa catggatatc tttgagaagg
     4021 ttgacgcaat ttttgacaag cacgaagttc aggagattaa ggagaagatc ctcaattccg
     4081 attatgacgt tgaggacttc ttcgaaggtg agttttttaa tttcgtgctc actcaagagg
     4141 gtatcgacgt gtataatgcg atcatcggtg ggttcgtgac tgagtccggt gaaaagatta
     4201 agggattgaa cgagtatatc aacctttaca accaaaagac gaaacagaag ctgccaaagt
     4261 tcaagcctct ttacaaacag gttctttcag accgcgagtc actctcgttc tatggggagg
     4321 gctacacttc ggatgaggaa gtcctggagg tgttcaggaa tactctcaat aagaattcgg
     4381 agattttctc ttctataaaa aaactggaaa agttgtttaa gaattttgac gaatactcta
     4441 gcgccggcat atttgtgaaa aacggcccgg ccatatcaac gataagtaaa gatatcttcg
     4501 gcgaatggaa cgtgatcaga gacaaatgga acgcggagta tgacgatatt cacctgaaga
     4561 agaaggctgt cgtaacggag aagtacgagg atgatcgcag gaaaagcttc aaaaagatcg
     4621 gaagtttcag cctggaacag ttgcaggagt atgctgacgc cgatcttagc gtcgtcgaga
     4681 agttgaagga gataatcatc caaaaggtcg acgagatata taaagtctat ggatcaagtg
     4741 aaaaactgtt cgacgccgac ttcgttttgg agaagtccct gaagaagaac gacgctgttg
     4801 ttgccattat gaaggatctg ctcgacagcg tgaagagttt cgagaactat attaaggctt
     4861 ttttcgggga ggggaaggag actaacagag atgagtcctt ctacggagac ttcgtcctcg
     4921 cgtacgatat actccttaag gtagaccaca tctacgacgc aatcagaaat tacgtgacac
     4981 aaaagccgta cagcaaggac aagttcaaac tctacttcca gaacccccag ttcatgggcg
     5041 gctgggacaa ggacaaggaa acggattaca gggctacgat cctgaggtat ggttcaaaat
     5101 actacttggc gattatggac aagaagtacg ccaagtgtct ccagaagatt gacaaagacg
     5161 atgtcaatgg caattatgag aagatcaact acaagctgct tccgggtccg aacaagatgc
     5221 tcccaaaggt tttcttcagc aagaaatgga tggcctacta taacccaagc gaggacatcc
     5281 agaagattta taagaacggt acgttcaaga agggcgacat gttcaatctt aacgactgtc
     5341 acaagctgat cgacttcttc aaagactcaa ttagccggta cccaaagtgg tctaacgcct
     5401 atgacttcaa cttttcggaa accgagaagt acaaggatat agccggattt tatagagagg
     5461 tggaagagca gggctacaag gtgtcattcg agtccgccag caagaaggaa gtggacaagc
     5521 tcgtggaaga gggtaagctc tacatgttcc agatttataa taaagacttt agcgataaga
     5581 gccacgggac acctaatctc cacacaatgt atttcaagct gctcttcgac gagaataacc
     5641 acggccaaat caggttgtca ggaggggctg aactcttcat gcggcgcgct agccttaaga
     5701 aggaggagct tgtagtccac cctgcgaata gtccaattgc gaataagaac ccggacaatc
     5761 ctaaaaagac tacaacattg agctacgacg tgtacaagga taagaggttt tccgaggatc
     5821 agtacgagct ccacatcccg attgcgatca acaagtgccc aaagaatatt ttcaagataa
     5881 acacagaggt gcgtgtactc ctgaagcatg acgacaatcc ttacgtcatt gggattgatc
     5941 ggggcgagag gaacctcctc tatattgtgg tggtggacgg gaaggggaac atagtcgaac
     6001 agtactccct taacgaaata attaacaatt tcaacggcat ccgtatcaag accgactacc
     6061 attcgttgct ggacaagaag gagaaggaga gatttgaggc gcggcaaaat tggacaagta
     6121 tcgagaacat caaggaactc aaagcaggtt atatctctca agttgtgcat aagatatgcg
     6181 agctggttga gaagtatgac gcagtgatcg ctcttgagga cctcaactcg ggctttaaga
     6241 attctagagt taaagtggag aagcaggtct atcaaaagtt cgagaagatg cttatagata
     6301 agctcaacta catggtcgat aagaaatcga acccatgtgc caccggcggc gcactcaaag
     6361 gttaccaaat aacaaacaaa ttcgagtcct tcaaatcgat gagtactcag aatgggttca
     6421 tattttatat accggcgtgg cttacgtcta agatcgaccc gtcaactggt tttgtcaacc
     6481 tgttgaagac gaaatacacg tccattgccg attcgaaaaa gttcatatct agttttgatc
     6541 gtattatgta cgtcccagag gaagatcttt tcgagtttgc tctcgactac aaaaactttt
     6601 cgcggaccga tgcggattac attaaaaaat ggaaactcta ttcgtacggc aacagaatca
     6661 ggatttttcg caaccctaag aagaataacg tctttgattg ggaggaagtt tgcttgacta
     6721 gcgcgtacaa ggagctcttt aataagtatg gcattaacta ccaacagggt gatatcagag
     6781 cactgctttg cgaacaatct gacaaggctt tctactcatc cttcatggct ttgatgagcc
     6841 tgatgctcca gatgagaaat tcaattacag gcagaaccga cgtggatttc ttgatctccc
     6901 cggttaaaaa ttctgatggc atcttttacg atagcaggaa ctatgaagcg caagagaatg
     6961 cgattctgcc aaaaaatgca gacgccaacg gtgcctataa catcgccagg aaagtcctgt
     7021 gggcgatcgg ccagttcaaa aaggccgaag acgaaaaatt ggacaaggtc aaaatcgcta
     7081 tcagcaacaa agagtggctg gagtatgctc agacatccgt aaagcataag cgtcctgctg
     7141 ccaccaaaaa ggccggacag gctaagaaaa agaagtgaga cgactagtgg cggccgccga
     7201 cgtccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc
     7261 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt
     7321 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt
     7381 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt
     7441 catctatgtt actagatcgg gaattgatcc cccctcgaca gcttccggaa agggcgaatt
     7501 cgcaactttg tatacaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
     7561 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     7621 actatgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     7681 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     7741 atcatcatgc ctcctc
//
