LOCUS       sequence-266604-        7008 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 7008)
  AUTHORS   Zhang Y, Ren Q, Tang X, Liu S, Malzahn AA, Zhou J, Wang J, Yin D, 
            Pan C, Yuan M, Huang L, Yang H, Zhao Y, Fang Q, Zheng X, Tian L, 
            Cheng Y, Le Y, McCoy B, Franklin L, Selengut JD, Mount SM, Que Q, 
            Zhang Y, Qi Y
  TITLE     Expanding the scope of plant genome engineering with Cas12a 
            orthologs and highly multiplexable editing systems.
  JOURNAL   Nat Commun. 2021 Mar 29;12(1):1944. doi: 10.1038/s41467-021-22330-w.
  PUBMED    33782402
REFERENCE   2  (bases 1 to 7008)
  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: "Nat 
            Commun."; date: "2021-03-29"; volume: "12(1):1944. doi"; pages: " 
            10.1038/s41467-021-22330-w"
COMMENT     Sequence Label: pYPQ284 (Mb2Cas12a)
FEATURES             Location/Qualifiers
     source          1..7008
                     /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             2585..2605
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6380..6427
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6458..6710
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6755..6878
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6895..6914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6896..6914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6919..6935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6919..6935)
                     /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 aaacatcatg
      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 aatactgtcc 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 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggctc tgtttcaaga
     2641 ttttacacat ctgtacccgc tgagtaaaac agtgcggttc gagctgaaac ccataggaag
     2701 gaccctcgag cacatccacg cgaagaattt tctgagccag gatgaaacta tggctgatat
     2761 gtatcaaaaa gttaaggtca ttttggacga ctatcatcgc gattttattg ccgacatgat
     2821 gggagaggtg aaactcacga agcttgctga attttacgac gtctatctga agttcaggaa
     2881 aaatcctaag gacgatgggc tgcaaaaaca gcttaaagac cttcaagctg tccttcggaa
     2941 ggaatcggtg aagcctatag ggtcaggtgg gaagtacaaa acaggctacg atagactctt
     3001 tggggcaaaa ctcttcaaag atggaaaaga gttgggtgac ctcgcaaaat tcgttatagc
     3061 ccaagaaggt gagtcttctc cgaagctggc tcatcttgct cattttgaga agttcagcac
     3121 gtattttact ggatttcacg ataatcggaa gaatatgtac tcggatgaag acaagcatac
     3181 tgcaatagcg tacaggctca tccatgagaa tttgccgaga ttcatcgaca atctgcaaat
     3241 cttgacaaca atcaaacaaa agcatagcgc cctctatgat cagataatca acgagctcac
     3301 ggcctccggg ctcgacgtct ccttggcttc tcatcttgac gggtatcaca agctccttac
     3361 acaagagggg atcacggcat acaacaggat cataggagag gtgaatggat atacaaataa
     3421 gcataaccag atatgccaca agagcgagcg catagcgaaa cttagaccct tgcacaagca
     3481 aatcctttct gacggaatgg gagtgtcatt ccttccgtct aagttcgcgg atgatagtga
     3541 gatgtgccaa gcggtcaacg aattttatcg ccattatact gacgtgttcg caaaggtgca
     3601 aagtctcttt gacggatttg atgatcacca gaaagacggg atctatgttg aacacaaaaa
     3661 ccttaatgaa ctgagcaaac aggcgttcgg cgactttgct ttgctgggga gggtccttga
     3721 tggatactac gtggacgttg tcaatccgga gttcaatgag cggttcgcaa aggccaagac
     3781 tgacaatgcg aaagccaagc ttacaaaaga aaaggacaaa ttcattaaag gagtccactc
     3841 actggcttcc ctcgaacaag caatagaaca ccatacagct agacacgacg atgagagtgt
     3901 tcaagccgga aaacttggcc agtacttcaa acacggtttg gcgggggttg acaacccgat
     3961 tcagaaaatt cacaataacc attcgacgat taaagggttt ctggaaaggg aaaggcctgc
     4021 tggggaacgg gcgctcccga agatcaagtc aggaaaaaac ccagaaatga cacagctcag
     4081 gcagctgaag gaacttttgg acaacgcatt gaatgtggcg cacttcgcta agctgctgac
     4141 aactaaaaca accttggaca accaggatgg aaatttttac ggggagtttg gggtgcttta
     4201 cgacgagctg gctaaaattc caactctcta caataaggtt agagattatc tctctcaaaa
     4261 gcccttttct accgaaaagt ataagctcaa cttcggcaat ccgacccttc tcaatgggtg
     4321 ggacctgaac aaagagaaag ataactttgg ggttatactt cagaaggatg gatgctatta
     4381 cttggcgctt cttgataagg ctcataaaaa agttttcgac aacgccccta acactggtaa
     4441 gaacgtctac caaaagatgg tctacaaact gttgcccggc cccaacaaaa tgcttcctaa
     4501 agtgtttttc gcaaaatcga atctcgacta ttataatcca tctgccgagc tccttgacaa
     4561 atatgctaag gggacccata aaaagggtga taatttcaac ctgaaggact gccacgcgct
     4621 tatcgacttt ttcaaagccg ggataaataa gcatccggag tggcaacatt ttggttttaa
     4681 attttcgcca acgtcgtcct atcgcgacct ttccgatttc tatagggaag ttgaacctca
     4741 ggggtaccag gtcaaatttg ttgacattaa tgcggactac attgatgaat tggtggagca
     4801 agggaagctc tacctctttc aaatatataa caaagatttc tcgccaaaag cgcatggtaa
     4861 accgaatctt cataccttgt actttaaagc acttttttca gaagataact tggcggaccc
     4921 gatctacaag ctgaatgggg aagctcagat cttctacagg aaagcttcgt tggacatgaa
     4981 cgagactacc atacatcgcg cgggagaggt gcttgagaac aaaaatcccg acaacccgaa
     5041 aaagcggcaa ttcgtttacg acatcatcaa agacaaacgg tacacgcagg acaaatttat
     5101 gctccacgtc cccattacca tgaattttgg agtccaaggc atgaccatta aggaattcaa
     5161 caaaaaggtc aaccaaagta ttcagcaata cgatgaagtc aatgtcatag gcatagatcg
     5221 gggagaaagg catctgttgt atcttaccgt gattaactct aagggtgaaa tactggagca
     5281 acggtcactt aacgatataa ccacggcgtc cgcgaacggt acacaagtga ccactcccta
     5341 ccacaaaata ttggataaaa gggagataga acgcttgaat gcccgcgttg gctggggtga
     5401 gattgagacc atcaaagagc ttaaatcggg atatttgtct cacgtcgttc atcaaattaa
     5461 ccaactcatg cttaagtaca atgcaatcgt tgtgctcgag gacctgaact ttggtttcaa
     5521 aagagggagg ttcaaggtgg aaaaacaaat ttaccagaac tttgaaaacg cgcttatcaa
     5581 gaaattgaat caccttgttt tgaaagataa ggcagatgac gaaatcgggt cgtataaaaa
     5641 tgcactccag ttgacaaata atttcacgga tttgaagtcg atcggcaagc aaacagggtt
     5701 cctcttttat gtgccagcgt ggaatacatc aaaaattgat ccggagacgg gatttgtcga
     5761 cttgctgaag cctaggtatg agaacattgc ccaatctcag gcctttttcg gcaaattcga
     5821 taaaatatgc tacaacacag acaaaggtta ttttgaattt cacattgatt acgccaaatt
     5881 tacagataag gcgaaaaaca gcagacagaa atgggctatc tgttctcatg gggacaaacg
     5941 ctatgtctac gataagacgg ctaatcaaaa taaaggcgcc gcaaaaggta ttaatgtgaa
     6001 tgatgagctg aaaagcttgt ttgcccgcta ccatatcaat gataaacaac caaacttggt
     6061 gatggacata tgccagaaca atgacaaaga attccacaag tcactcatgt gcctgcttaa
     6121 aacccttttg gcgctgcggt atagcaatgc atctagcgat gaagacttta ttttgagtcc
     6181 cgtggccaac gacgagggcg tgttttttaa ttcagccttg gcggacgata cgcagcccca
     6241 gaatgcggac gcaaacggcg cgtaccacat tgcactgaag ggactgtggc ttctgaacga
     6301 gctgaaaaat agcgacgacc tgaataaagt caagttggcc attgacaatc aaacctggtt
     6361 gaatttcgct caaaatagaa agcgtcctgc tgccaccaaa aaggccggac aggctaagaa
     6421 aaagaagtga gacgactagt ggcggccgcc gacgtccgat cgttcaaaca tttggcaata
     6481 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     6541 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     6601 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     6661 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattgat
     6721 cccccctcga cagcttccgg aaagggcgaa ttcgcaactt tgtatacaaa agttgaacga
     6781 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     6841 acataatact gtaaaacaca acatatccag tcactatgcc atccagctga tatcccctat
     6901 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     6961 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctc
//
