LOCUS       sequence-272077-        6873 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 6873)
  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 6873)
  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: pYPQ285 (Lb5Cas12a)
FEATURES             Location/Qualifiers
     source          1..6873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     816..838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     830..847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             987..1007
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4647..4694
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4725..4977
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    5022..5145
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5162..5181)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5163..5181)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5186..5202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5186..5202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5633..6424
                     /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      join(6517..6873,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgcgagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc ttgcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
      961 ctccgaattc gcccttcacc atggctccta agaagaagcg gaaggttggt attcacgggg
     1021 tgcctgcggc ttactatgaa tccctgacca aacaataccc tgtttccaaa acaatccgca
     1081 atgagctgat tcctataggt aaaaccctgg ataacatccg ccaaaataac atcttggaat
     1141 cagacgttaa aagaaagcag aattacgaac atgtcaaagg tatacttgat gaataccaca
     1201 aacagctcat aaacgaagcg ttggataact gtacgcttcc ctcactgaaa atagcggctg
     1261 agatctacct caaaaaccag aaggaggtgt cagacagaga agatttcaac aaaacccagg
     1321 atctgttgcg caaggaggtt gtggagaaac tcaaggcgca tgaaaatttt actaagatag
     1381 gaaaaaaaga tattcttgat ttgttggaga agcttcctag catatccgag gacgactaca
     1441 atgccctgga aagtttcaga aacttttaca catacttcac atcgtataat aaggtccggg
     1501 aaaatctgta tagcgataaa gagaaaagtt ctactgttgc gtacaggctt atcaatgaga
     1561 attttccaaa gtttctcgac aacgtcaaat catatcggtt cgtcaaaact gcgggcattt
     1621 tggctgacgg attgggagag gaggagcagg acagcctgtt catagtggag actttcaaca
     1681 agacattgac ccaggatggc attgatacat ataactccca ggtcggcaag ataaactcct
     1741 cgataaacct ctacaaccag aaaaaccaga aggcaaacgg cttccggaaa atcccaaaga
     1801 tgaaaatgct ttataagcag atcttgagtg atcgggagga gtctttcatc gatgaatttc
     1861 agtcagacga agttcttatc gacaacgttg agagttacgg ctctgtgctt attgagagcc
     1921 tcaagtcgtc aaaggtttct gcattttttg atgcacttcg ggagagtaaa ggtaaaaacg
     1981 tttacgttaa gaatgacctg gcgaaaacag caatgtcaaa catagttttt gagaactgga
     2041 ggaccttcga tgaccttctg aatcaagagt acgatttggc gaacgaaaat aaaaagaagg
     2101 acgacaagta ctttgagaag aggcaaaagg agctgaaaaa gaataaatcg tattcgttgg
     2161 aacatctttg caacctctct gaggattcct gcaacctgat agaaaactac atccaccaga
     2221 tcagtgacga tattgaaaac attattatta acaatgaaac ctttttgcgg atagtgatta
     2281 atgagcacga tcgcagtaga aaacttgcta aaaatagaaa agctgttaaa gcaataaagg
     2341 atttccttga cagtattaag gtgctcgagc gggagttgaa actgatcaat tcttcaggac
     2401 aagagttgga aaaagacctt atcgtctata gcgctcacga ggaacttctg gtggaactga
     2461 aacaagttga ttcgctttat aacatgacga ggaactacct gaccaaaaag cccttttcta
     2521 ctgagaaagt caaactcaat tttaatcggt cgacgcttct gaatggctgg gaccgcaaca
     2581 aagagactga taacctcggg gttctcttgc tgaaagatgg caagtattat ctgggtataa
     2641 tgaatacatc agcaaataag gcattcgtta acccgcctgt ggctaaaacc gagaaggttt
     2701 ttaaaaaagt ggactacaag ctgttgccag tgccgaacca gatgttgccc aaggtttttt
     2761 ttgcgaaaag caatattgat ttctataatc catcgagcga gatatattcc aactacaaaa
     2821 aaggtaccca taaaaagggt aacatgtttt cacttgagga ctgccacaac ctgattgatt
     2881 tctttaaaga gagtattagt aagcacgaag attggagtaa gtttggattc aaattttctg
     2941 atactgcgag ttacaatgat ataagtgaat tctatcggga agtcgaaaaa cagggttaca
     3001 agcttacgta cactgacata gatgaaacat acattaatga cctgattgag cgcaatgagt
     3061 tgtacctgtt tcagatctat aacaaagatt tttccatgta tagcaaaggc aagctcaact
     3121 tgcataccct gtattttatg atgctcttcg atcaaaggaa tatcgatgac gttgtttata
     3181 agctgaatgg tgaagcagaa gttttttacc gcccagcctc gatcagtgaa gatgagctga
     3241 tcattcacaa agcgggtgaa gagattaaga ataagaaccc taatcgggcc aggacgaaag
     3301 aaacttcaac gttttcctat gatattgtca aagacaaaag gtactcgaag gataagttta
     3361 cccttcacat tcccataacc atgaattttg gtgtcgacga agtcaagcgg tttaacgacg
     3421 ccgtgaattc cgcgatccgg atagacgaga atgtgaatgt cattgggatt gacagaggtg
     3481 aacgcaatct tctttacgtg gtggtgatag actctaaagg gaatatattg gagcaaattt
     3541 ctctgaactc aatcattaat aaagaatacg atattgaaac agactatcat gcccttctcg
     3601 acgagcggga gggcgggagg gataaagcac ggaaggactg gaacacagtt gagaatatcc
     3661 gggacctcaa agccggatac ctttcccagg tggtcaatgt tgtcgcaaaa ttggtcttga
     3721 agtacaatgc cattatatgc cttgaagatt tgaactttgg gttcaaacgc ggtaggcaaa
     3781 aagttgaaaa acaagtttac cagaaattcg aaaagatgct tatagataag ctcaactacc
     3841 tggttattga caagagtaga gagcagacgt cgcccaaaga actcgggggg gcgctgaatg
     3901 cgttgcaact gacctcgaag ttcaaatcgt tcaaagaatt ggggaaacag tccggcgtta
     3961 tctattacgt gcctgcttat ctcacgtcta agatagatcc taccacaggc ttcgcgaatc
     4021 ttttttatat gaagtgtgag aatgtcgaga agtctaagcg gtttttcgat ggatttgact
     4081 tcatcaggtt caatgcgctc gaaaatgttt ttgagtttgg attcgactac cggagcttta
     4141 cacagagagc gtgcggtata aacagtaaat ggacagtctg cacaaatggg gagagaatta
     4201 taaagtatag gaaccccgat aaaaataaca tgttcgacga aaaggtggtg gttgttactg
     4261 atgaaatgaa gaacctgttt gaacagtaca aaatacctta tgaggatgga aggaacgtga
     4321 aggacatgat tatttcaaac gaggaggcgg agttctaccg cagattgtac cgcctgcttc
     4381 agcagaccct ccaaatgcgg aattcgacct cagatggaac gagagattat attatatcac
     4441 cagtgaaaaa caagcgcgag gcgtatttca actctgagct ttccgatggt tcggttccga
     4501 aagacgcaga tgcaaacgga gcatacaaca tcgcaaggaa agggttgtgg gtgttggaac
     4561 aaatccgcca gaaatccgaa ggcgaaaaaa ttaacttggc tatgaccaat gctgagtggc
     4621 ttgaatacgc gcagacgcac cttctgaagc gtcctgctgc caccaaaaag gccggacagg
     4681 ctaagaaaaa gaagtgagac gactagtggc ggccgccgac gtccgatcgt tcaaacattt
     4741 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     4801 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     4861 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     4921 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg
     4981 aattgatccc ccctcgacag cttccggaaa gggcgaattc gcaactttgt atacaaaagt
     5041 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
     5101 acagactaca taatactgta aaacacaaca tatccagtca ctatgccatc cagctgatat
     5161 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     5221 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctggac
     5281 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     5341 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     5401 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     5461 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     5521 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     5581 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     5641 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     5701 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     5761 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     5821 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     5881 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     5941 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     6001 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     6061 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     6121 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     6181 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     6241 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     6301 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     6361 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     6421 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     6481 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6541 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6601 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6661 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6721 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6781 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6841 gggttcgtgc acacagccca gcttggagcg aac
//
