LOCUS       sequence-266613-        7041 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 7041)
  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 7041)
  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: pYPQ288 (BoCas12a)
FEATURES             Location/Qualifiers
     source          1..7041
                     /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             6413..6460
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6491..6743
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6788..6911
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6928..6947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6929..6947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6952..6968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6952..6968)
                     /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 ggaagtttaa
     2641 tgagttcgtt ggcctttacc caattagtaa aacattgaga tttgaattga agcccatagg
     2701 gaagacactc gaacatatcc agcggaacaa actgcttgag cacgatgcgg tccgggctga
     2761 cgattacgtg aaggttaaga agataatcga caaatatcac aagtgtctca tcgatgaagc
     2821 gttgtcgggg ttcacatttg atactgaggc cgacgggagg tctaataaca gcttgtccga
     2881 gtattatctt tattacaatc tgaaaaagcg gaacgaacag gagcagaaaa cgttcaaaac
     2941 catacaaaat aatttgagga aacaaatagt caataagttg acgcaaagcg agaagtacaa
     3001 acggatcgac aagaaagaac tgatcactac tgacctcccc gatttcctca ctaatgagtc
     3061 ggaaaaggaa ttggttgaaa aatttaagaa ctttacaact tacttcacgg agttccacaa
     3121 gaacaggaaa aatatgtatt ctaaagagga aaagtctaca gcgatagcct tcaggcttat
     3181 aaatgaaaat ttgcccaagt tcgttgacaa tatagctgcg tttgaaaaag tcgtgtcgag
     3241 cccattggcg gaaaaaatta atgctctcta cgaggatttt aaggaatacc ttaacgttga
     3301 agagatttct agggttttta gacttgacta ctatgacgaa ctgctgactc agaagcagat
     3361 tgacctttat aacgctatag ttggcggcag gaccgaggag gataataaaa ttcagattaa
     3421 aggactcaac caatatatca acgaatacaa tcagcagcag acagatagat cgaacagact
     3481 cccgaaactt aagccactgt ataaacagat tctttccgat agggaatccg tctcctggct
     3541 gcctccgaaa tttgactcag acaaaaacct cctcataaag attaaagaat gttacgacgc
     3601 gctctctgag aaagaaaaag tttttgataa actggaaagt attcttaaaa gtctctcaac
     3661 ttacgatttg tctaagatat atatatcaaa cgacagccaa ttgagctaca tcagccaaaa
     3721 gatgttcggg aggtgggaca taataagtaa agcaattcgg gaagactgcg ctaaaaggaa
     3781 cccgcaaaag tcaagggaga gtttggagaa gtttgctgag cgcatagata aaaaattgaa
     3841 gacgatagac agtatctcga ttggggatgt tgacgagtgt ctggcgcaac tcggtgaaac
     3901 atacgtgaaa cgggttgagg actactttgt cgctatgggt gagtcagaga tagacgacga
     3961 acaaacggac acaacttctt ttaaaaaaaa tatagaagga gcgtatgaat ccgtcaagga
     4021 acttctcaat aacgcggaca acattactga taacaatctg atgcaagata aaggaaatgt
     4081 ggaaaagatt aaaacgttgc ttgacgctat aaaggacctc cagcgcttta taaaacccct
     4141 tctcggcaag ggtgatgagg cggacaaaga tggtgtcttt tatggggagt tcaccagtct
     4201 ttggactaaa ttggatcaag tcaccccact ctataacatg gtgaggaatt accttacgtc
     4261 taagccgtat agtaccaaaa aaattaaact taactttgaa aactcgaccc tcatggacgg
     4321 ctgggacctc aataaagaac ccgataacac tacagttatt ttctgcaaag acggcttgta
     4381 ttacctcggc attatgggca agaagtacaa cagggttttc gtcgatagag aggacctccc
     4441 gcacgacggt gaatgctacg acaagatgga gtataagctg cttccgggag cgaacaagat
     4501 gcttcctaag gtgttctttt ctgaaacggg cattcaaaga tttcttccgt ccgaggagtt
     4561 gttgggtaaa tacgagaggg ggacgcataa gaaaggagca ggatttgact tgggagattg
     4621 tcgggcgctt atcgattttt tcaaaaagag tattgagcgc cacgatgatt ggaagaaatt
     4681 cgactttaag ttctctgata ccagcaccta ccaggacatt tcagagttct acagagaggt
     4741 cgagcaacag ggatataaaa tgtcattcag aaaggttagt gtcgattata tcaagtcact
     4801 cgttgaagaa ggtaaactct atcttttcca gatttacaat aaggatttct ctgctcatag
     4861 taaggggacc ccgaatatgc acaccttgta ctggaaaatg ctgtttgacg aagaaaatct
     4921 taaggacgtc gtttacaagc tgaatggaga ggcggaagtc ttttttagga aatcaagtat
     4981 tacggtccaa tcgcccacgc atccggcgaa ttctcctata aagaataaga ataaggataa
     5041 tcaaaaaaaa gaaagtaagt tcgagtacga tctgattaaa gacagacggt atactgttga
     5101 taagttcctg tttcatgttc ccattactat gaacttcaaa agcgtcggtg gttcgaatat
     5161 taaccaattg gttaagcgcc atatacgcag cgctaccgac ttgcacataa tcggaatcga
     5221 ccggggtgag aggcacttgc tttatcttac cgttatagac tctaggggta acataaaaga
     5281 acaattctct cttaacgaga tagttaatga atataacgga aatacttacc gcacagatta
     5341 ccatgagctc ctcgacacgc gcgaggggga gcggacggag gctcgcagga actggcagac
     5401 tatacaaaat ataagggaac tgaaggaagg ctatttgtcc caggtgatac acaagatttc
     5461 ggagttggcg attaagtata atgcagtcat tgtgctggag gatctgaatt ttgggtttat
     5521 gaggtcgcgg cagaaagttg aaaaacaagt ctaccagaag ttcgagaaga tgctcataga
     5581 caaactgaat tacttggtcg ataagaaaaa gccggttgct gaaaccggag ggctcctcag
     5641 ggcgtaccag ctcaccggcg aatttgagag ctttaaaacg ctcggtaagc aatctgggat
     5701 cttgttctac gtcccagctt ggaatacctc gaaaatcgat ccagtgacgg gctttgtgaa
     5761 tctttttgat acgcactacg agaatataga gaaagcaaaa gttttcttcg ataaatttaa
     5821 gtcgatccgg tataacagtg ataaggactg gtttgaattc gtcgtcgatg actacacaag
     5881 atttagtccc aaagccgaag ggactagacg cgattggacc atatgcacac aggggaagag
     5941 gatacaaatt tgtcgcaatc accagcggaa taatgaatgg gaaggtcaag agattgatct
     6001 gacaaaagcg tttaaggaac attttgaggc ctatggcgtg gacatctcaa aggacttgcg
     6061 cgaacagatt aacacgcaga ataaaaaaga gttttttgaa gaattgctcc ggctgctcag
     6121 gctcacgttg caaatgagga actctatgcc atctagcgat attgattacc tcatatctcc
     6181 agttgccaat gataccggtt gcttcttcga tagtagaaaa caggcagaac tgaaagagaa
     6241 cgctgtcttg cctatgaatg cagacgcaaa tggtgcttac aatattgcac gcaaggggct
     6301 cctcgcaatc agaaagatga agcaagagga gaatgactca gcgaaaattt ctttggcaat
     6361 tagtaataaa gaatggctga aattcgctca gactaagcct tacttggaag ataagcgtcc
     6421 tgctgccacc aaaaaggccg gacaggctaa gaaaaagaag tgagacgact agtggcggcc
     6481 gccgacgtcc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
     6541 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
     6601 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
     6661 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
     6721 ggtgtcatct atgttactag atcgggaatt gatcccccct cgacagcttc cggaaagggc
     6781 gaattcgcaa ctttgtatac aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat
     6841 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     6901 cagtcactat gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     6961 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     7021 aatatatcat catgcctcct c
//
