LOCUS       sequence-272045-        6948 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 6948)
  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 6948)
  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: pYPQ239-STU
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..41
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     33..50
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     34..50
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    66..165
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             190..210
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4132..4179
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    4300..4423
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4440..4459)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4441..4459)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4464..4480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4464..4480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4911..5702
                     /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      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6284..6303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6537..6554
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6713..6740)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6832..6918)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgcttgcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
       61 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      121 agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccgaa ttcgcccttc
      181 accatggctc ctaagaagaa gcggaaggtt ggtattcacg gggtgcctgc ggcttcgatc
      241 tatcaagaat tcgttaataa gtattcgctt tctaaaacac tgagattcga actcatacct
      301 caaggaaaga cactcgagaa cataaaggct aggggcctca tattggatga tgaaaagaga
      361 gccaaggact ataaaaaagc caaacagata atcgacaagt accaccaatt cttcatagaa
      421 gaaatccttt ccagcgtctg cattagcgag gatttgttgc aaaattacag cgatgtctac
      481 ttcaagctta agaagagcga tgacgacaat ctccagaaag acttcaaatc agcgaaggac
      541 acaattaaga agcagatcag cgagtatatc aaagatagtg aaaaattcaa gaaccttttc
      601 aatcagaacc tgatagacgc aaaaaaagga caagaaagcg acctcattct ttggttgaag
      661 cagtctaagg acaacgggat tgagcttttt aaggcgaata gcgatataac cgacatcgac
      721 gaggcgcttg agatcataaa gtcgtttaag ggatggacaa catacttcaa aggcttccat
      781 gagaatcgca aaaacgtcta ctccagcaac gacattccaa cgtcgattat atacagaatt
      841 gttgatgata acctccctaa attcctcgaa aataaggcaa aatatgaaag ccttaaagat
      901 aaagcgcctg aagcaatcaa ttatgaacaa atcaaaaagg atcttgctga agaattgacg
      961 tttgatatag actacaagac gtcagaagtt aaccagaggg tgttttcact cgacgaggtg
     1021 tttgagattg ctaatttcaa caactacctc aatcagagtg ggatcacgaa gttcaacact
     1081 attataggtg gtaagttcgt gaatggtgag aacactaaaa gaaaagggat taacgaatat
     1141 ataaaccttt atagtcaaca gatcaacgac aagactttga agaaatataa gatgagcgtc
     1201 ctcttcaagc agatactcag tgacacggaa tccaagagct ttgtgatcga caagctcgaa
     1261 gatgactcgg atgtggtcac taccatgcaa tccttctacg agcaaattgc cgctttcaaa
     1321 actgtggagg agaagagtat aaaggagaca ctgtctctgc tttttgacga tcttaaggcc
     1381 cagaaattgg atctttccaa aatatatttc aagaatgata agtcacttac ggacctttcc
     1441 caacaagttt ttgacgatta ttcagttatt ggtacggcgg ttcttgagta cattacgcag
     1501 cagatagccc ccaagaatct ggacaacccc tctaagaaag aacaggagtt gatagcgaag
     1561 aagacagaga aggcgaaata cctctcgctg gagaccataa aattggcact ggaagaattt
     1621 aacaagcatc gcgacataga taaacagtgc cgcttcgagg aaattttggc aaattttgca
     1681 gccattccaa tgattttcga cgagatagcg caaaacaagg ataatttggc acaaatttca
     1741 ataaaatatc aaaaccaggg caagaaggac ctcttgcagg cttcggcaga ggatgatgtt
     1801 aaggctatta aagacttgtt ggaccaaacg aacaatctgt tgcacaagtt gaaaattttc
     1861 cacattagtc aaagcgagga taaagcaaac atattggaca aagacgagca cttttatctg
     1921 gtgtttgagg agtgctactt cgagcttgct aatattgtcc cactttataa taaaataaga
     1981 aactacatta cgcaaaagcc atattcagat gaaaagttta agctcaattt cgaaaatagt
     2041 actcttgcca acggctggga caagaataag gagccagata ataccgccat actttttatc
     2101 aaagatgata aatattatct tggggtgatg aataagaaga ataataagat cttcgatgat
     2161 aaagcgataa aggaaaataa gggtgaaggc tataaaaaaa ttgtttacaa actgttgccg
     2221 ggagcaaata aaatgctccc caaggttttt ttttcggcaa agagcattaa attttacaat
     2281 ccttcagaag acattctgcg cataagaaat cattcgacac acactaaaaa tggttcgcca
     2341 caaaagggct acgagaaatt tgaattcaac attgaggact gtcggaagtt cattgatttc
     2401 tacaagcagt ccatctccaa gcacccggag tggaaagatt ttgggtttcg gttttccgac
     2461 acgcagagat acaacagcat tgatgaattt tatagagagg tcgagaatca aggttataag
     2521 cttacctttg aaaacatttc tgaatcatac attgattcag tggtcaatca gggcaaactc
     2581 tatctttttc aaatatacaa caaggacttt agtgcttata gtaaagggcg gcccaatttg
     2641 catactctct attggaaagc gctgtttgat gagcggaacc ttcaagacgt cgtgtataag
     2701 ctcaacgggg aagccgagct cttttaccgc aagcagtcca taccgaaaaa aataacacac
     2761 cctgccaaag aagccatcgc caacaagaat aaagacaatc ctaaaaaaga gtccgtcttc
     2821 gaatatgatc ttattaagga caagaggttt acagaagata aatttttctt ccattgtccc
     2881 ataactatca atttcaaaag ctctggcgcg aacaaattta acgacgaaat caatctcttg
     2941 ttgaaagaaa aagccaacga tgtgcacatt ctgtcgatcg acaggggaga gcgccacttg
     3001 gcatactaca cccttgttga tgggaaagga aatattatta aacaggacac atttaatatc
     3061 atcggcaacg atcgcatgaa gaccaactat catgacaaac tggcagcaat tgaaaaggac
     3121 cgcgactcag cgagaaaaga ctggaagaag atcaataata tcaaagaaat gaaagagggt
     3181 tatttgtctc aagtggtcca tgagatcgcg aagttggtca ttgaatataa tgccatagtg
     3241 gtcttcgaag atctgaattt tggatttaag cgcggcaggt tcaaagtcga aaaacaggtc
     3301 taccaaaagt tggaaaagat gctcatcgaa aagctgaatt accttgtctt caaagataac
     3361 gaattcgata aaaccggggg ggtcttgagg gcctaccaac tgactgcacc ctttgagact
     3421 tttaaaaaga tgggtaaaca gacaggaata atttactatg ttcctgccgg tttcactagc
     3481 aagatttgcc ccgttaccgg attcgtgaat caactctatc ccaaatacga atccgtgagc
     3541 aagagtcagg aattcttctc caaatttgat aaaatatgct ataatctcga caaaggttat
     3601 ttcgagttct cgttcgacta taagaacttc ggggataagg ctgccaaggg aaagtggact
     3661 atagcaagct ttggtagtcg ccttataaat tttaggaaca gcgacaagaa tcacaactgg
     3721 gacactcggg aagtctaccc aacaaaagaa ctggagaaac tcttgaagga ttatagtatc
     3781 gagtatgggc atggggagtg tatcaaggca gcgatttgtg gagagtccga caaaaagttt
     3841 tttgctaaac tcacctcggt gctcaacact atcctccaga tgagaaattc aaaaacaggg
     3901 acagagctcg attacctcat tagccccgtt gccgacgtca atggaaactt tttcgactca
     3961 agacaggctc caaaaaacat gccgcaagat gcggacgcga atggggccta tcacataggc
     4021 ctgaaagggc ttatgctcct tgggagaatt aaaaataacc aagaaggcaa aaaactcaac
     4081 ctcgtcatta agaacgaaga atacttcgaa tttgttcaga acaggaataa caagcgtcct
     4141 gctgccacca aaaaggccgg acaggctaag aaaaagaagt gagacgacta gtggcggccg
     4201 ccramrwccw aaaaaaaaaa aaaaaraamm aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4261 aaaaaaaaaa aaaaaaaaaa tccggaaagg gcgaattcgc aactttgtat acaaaagttg
     4321 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     4381 agactacata atactgtaaa acacaacata tccagtcact atgccatcca gctgatatcc
     4441 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     4501 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgcctc ctctggacca
     4561 gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt
     4621 ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat
     4681 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     4741 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc
     4801 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
     4861 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag
     4921 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc
     4981 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agttgatggc ggcctgaagc
     5041 cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc
     5101 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc
     5161 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta
     5221 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc
     5281 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg
     5341 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg
     5401 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc
     5461 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc
     5521 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca
     5581 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag
     5641 atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat
     5701 aaccctcgag ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6541 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6601 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6661 gcaacgcaat taatacgcgt accgcgagcc aggaagagtt tgtagaaacg caaaaaggcc
     6721 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     6781 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     6841 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     6901 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaa
//
