LOCUS       sequence-272081-        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: pYPQ287 (BsCas12a)
FEATURES             Location/Qualifiers
     source          1..6873
                     /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             6245..6292
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6323..6575
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6620..6743
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6760..6779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6761..6779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6784..6800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6784..6800)
                     /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 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggctt actatcagaa
     2641 tctcactaag aagtacccag tcagcaaaac tatcagaaat gaacttattc caataggaaa
     2701 aacgcttgaa aacattagga aaaacaacat actggaaagc gacgtgaaaa ggaagcagga
     2761 ctatgaacac gttaagggga tcatggacga atatcacaag cagctcatta acgaagcact
     2821 tgataactat atgcttcctt cgctcaatca agccgccgaa atttacctca agaaacacgt
     2881 ggacgtcgag gatagagagg agtttaagaa aacgcaagat ctgctcaggc gggaggtgac
     2941 gggaagactg aaggaacatg agaattacac taagattgga aaaaaggaca ttctcgatct
     3001 cctcgaaaaa ctgccatcta tttctgagga agactacaat gctctggagt cctttagaaa
     3061 cttttataca tatttcacct cctataataa agttcgggag aatctgtata gcgacgaaga
     3121 gaagagtagc acggtggcct atagactgat aaacgagaac ttgccaaagt tcttggataa
     3181 catcaaatct tatgcatttg ttaaagcggc tggcgtgctc gctgattgta ttgaagagga
     3241 ggaacaggat gccctgttta tggtcgaaac tttcaacatg acgctcacgc aagaaggtat
     3301 tgacatgtat aattatcaga tcggaaaagt caattcggct attaatctct ataaccagaa
     3361 gaatcacaag gtcgaggaat ttaagaaaat cccaaaaatg aaagttcttt ataaacagat
     3421 cctctctgat cgcgaagaag tcttcatcgg tgaattcaag gatgatgaga ctctcctttc
     3481 cagcataggc gcatatggca atgttttgat gacatacctc aaatccgaga aaataaatat
     3541 tttcttcgac gctctgaggg aaagcgaggg aaaaaacgtc tacgttaaga acgatctgtc
     3601 gaaaactacg atgtcaaata tagtctttgg ctcgtggtcg gcctttgatg aacttctgaa
     3661 tcaggaatac gacctggcca atgagaacaa aaagaaagat gataaatact ttgagaagag
     3721 acagaaggaa ctcaaaaaaa ataagagcta cacgttggag caaatgagca atctttcgaa
     3781 ggaggatatt tcgccgatag agaactacat agagaggatt tcagaggaca tagagaagat
     3841 atgcatctat aatggcgagt tcgaaaagat tgtcgtgaac gaacatgatt cgagtagaaa
     3901 gctctcgaag aacatcaagg cggttaaggt gattaaagac tacctcgatt ccattaaaga
     3961 acttgaacat gatattaaac tcataaatgg atctggtcag gagcttgaaa agaacttggt
     4021 tgtttacgtc gggcaggaag aagcgcttga acagctgcgc cccgtcgatt ccctgtataa
     4081 cctcacacgc aattatctga cgaagaagcc attttcaacg gaaaaagtga aactgaattt
     4141 taataaatct actctcctca atggctggga caaaaataag gaaacggata acctcggtat
     4201 actgtttttt aaggacggta aatactacct cggaataatg aacaccacag caaacaaggc
     4261 gtttgtcaac ccacctgctg ccaaaacaga gaatgttttc aaaaaggtgg actacaagct
     4321 gctgcctggc tcgaacaaaa tgctcccaaa ggttttcttt gcgaaatcga atattggata
     4381 ttataatccc tcgaccgaac tctattccaa ctacaaaaaa ggcactcaca agaaaggccc
     4441 ttccttctct atagatgatt gccacaatct tattgatttt tttaaagaat cgattaagaa
     4501 gcatgaagac tggtccaaat ttggatttga attcagtgac actgcagatt acagagacat
     4561 ttcggagttt taccgggaag tggagaagca ggggtataag ttgactttca ccgacataga
     4621 tgagtcatac atcaacgatc tgatagaaaa gaacgaactg tatctttttc agatttataa
     4681 caaagatttc tctgaatatt cgaagggtaa gctgaacctc cacacactct attttatgat
     4741 gctgttcgac cagagaaacc ttgacaacgt cgtgtataag ctgaatggag aggccgaggt
     4801 cttctataga ccagcatcta tagccgaaaa cgaacttgtg atacacaaag cgggcgaagg
     4861 cataaagaac aaaaacccta atcgggcgaa agttaaagag actagtacat tcagttacga
     4921 cattgtcaaa gataagcggt attcaaagta caagttcaca ctgcatattc ctatcacaat
     4981 gaatttcgga gttgacgagg tccgcagatt taacgacgtt atcaataacg cactgcgcac
     5041 agacgataac gtgaatgtta tcggtatcga ccggggcgaa cgcaatctgt tgtatgtcgt
     5101 ggtgatcaat tcagagggta agatcttgga gcaaatttcc ttgaatagta taattaacaa
     5161 agagtatgac atagagacta attatcatgc cctgttggac gaacgcgaag acgatagaaa
     5221 taaagcaaga aaagattgga acactatcga aaacattaaa gaattgaaaa caggttatct
     5281 gagtcaagtc gtcaacgttg tcgccaaact cgtcctgaaa tataacgcga taatctgctt
     5341 ggaagacctt aattttggtt tcaagcgcgg acggcagaag gtcgaaaaac aggtctatca
     5401 aaaatttgag aaaatgctca tagagaaact taattatttg gttattgata agagtagaga
     5461 gcaggtgtcc cccgaaaaga tggggggtgc cctcaacgct ctccaattga cttcaaaatt
     5521 taagagcttt gcggaacttg ggaagcagag tggtatcatt tactatgttc ccgcgtactt
     5581 gacttctaag attgatccta ccacgggttt tgtgaatctg ttctacatca agtatgagaa
     5641 catagaaaag gcaaagcaat ttttcgatgg ttttgatttc attcggttta ataagaaaga
     5701 cgacatgttt gaatttagct ttgactacaa gtcattcacc cagaaggcgt gcgggatacg
     5761 gtcgaaatgg atagtctaca caaatggaga gcgcatcata aaatatccga accctgagaa
     5821 gaataacctg ttcgacgaaa aggttattaa cgtcacggac gagattaaag gtctttttaa
     5881 gcaataccgg attccgtacg agaacggaga agacattaag gaaataatca tttcgaaggc
     5941 agaggctgac ttctataaac ggttgttccg cctgcttcat caaacgctgc aaatgagaaa
     6001 ttcaactagc gatggcacca gggattacat catcagtccg gtgaagaacg accggggaga
     6061 atttttctgc tctgaatttt cggagggcac aatgcctaaa gatgccgacg cgaacggtgc
     6121 atacaatata gccagaaagg ggctttgggt cctcgagcag attcgccaga aagatgaggg
     6181 agagaaagtt aatttgtcaa tgacgaatgc cgaatggctt aaatacgccc aattgcacct
     6241 cctgaagcgt cctgctgcca ccaaaaaggc cggacaggct aagaaaaaga agtgagacga
     6301 ctagtggcgg ccgccgacgt ccgatcgttc aaacatttgg caataaagtt tcttaagatt
     6361 gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca
     6421 tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt
     6481 cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa
     6541 attatcgcgc gcggtgtcat ctatgttact agatcgggaa ttgatccccc ctcgacagct
     6601 tccggaaagg gcgaattcgc aactttgtat acaaaagttg aacgagaaac gtaaaatgat
     6661 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     6721 acacaacata tccagtcact atgccatcca gctgatatcc cctatagtga gtcgtattac
     6781 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     6841 gcacaagata aaaatatatc atcatgcctc ctc
//
