LOCUS       sequence-272080-        6936 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 6936)
  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 6936)
  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: pYPQ286 (CMaCas12a)
FEATURES             Location/Qualifiers
     source          1..6936
                     /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             6308..6355
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6386..6638
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6683..6806
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6823..6842)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6824..6842)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6847..6863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6847..6863)
                     /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 cctgcggctg acgcgaaaga
     2641 gtttactgga caatacccac tctcaaagac tctccgcttt gagctgagac ccatcgggag
     2701 gacttgggac aacttggaag ccagtggcta cttggccgag gaccgccacc gcgccgagtg
     2761 ctacccgaga gcaaaagagc tgctggatga caaccataga gcgttcctga acagggttct
     2821 cccgcagatt gatatggatt ggcatcccat cgcagaagcc ttctgcaaag tccataaaaa
     2881 tccaggaaac aaagagctcg cgcaagatta taatcttcag ttgtccaaga gacgcaaaga
     2941 aatctcggct tatcttcagg acgctgacgg atataaaggt ttgtttgcta aacctgctct
     3001 tgacgaggca atgaagatcg ctaaggagaa cggtaacgaa tcggatattg aggtcctcga
     3061 ggcatttaac ggcttttccg tttatttcac gggctaccac gagtctagag aaaatatcta
     3121 ctcagacgaa gacatggtct ccgtggccta caggatcaca gaggataatt ttccccgctt
     3181 cgtctccaac gcgttgatat tcgacaagtt gaatgaatca catcctgata tcatctctga
     3241 ggtgtctggg aatctcggtg ttgacgatat aggcaaatac tttgacgtct cgaactacaa
     3301 taactttctc agccaggcag gaattgatga ctataatcat attattggtg gacacaccac
     3361 tgaagatggt ctgattcagg cctttaatgt ggttttgaat ctgcgccacc agaaggatcc
     3421 agggttcgaa aagatccaat tcaaacagct ctataagcag attctctctg ttcggacgtc
     3481 caaatcgtat attccaaaac aattcgacaa ttctaaggaa atggtcgatt gcatttgcga
     3541 ttatgtctca aaaatcgaga aatcagagac ggtcgaaagg gctttgaagc ttgtgagaaa
     3601 tatctccagc ttcgatcttc ggggcatatt tgtgaacaag aaaaacctca ggatcctctc
     3661 caataagttg ataggggatt gggacgcaat cgagaccgcg cttatgcatt ccagctcctc
     3721 agaaaatgat aaaaaatccg tctatgattc agccgaagcc tttactttgg acgatatatt
     3781 ctcgtcagtg aaaaaattca gtgatgctag cgccgaggat attgggaacc gcgccgagga
     3841 catatgcagg gtgatctctg aaacggctcc ctttattaac gacttgcgcg cggtggacct
     3901 ggacagtctc aacgatgacg gatatgaagc agcagttagc aagatccggg agtcccttga
     3961 gccgtacatg gatcttttcc atgaacttga gatatttagt gttggggacg agttccctaa
     4021 atgtgcggca ttttactcgg agttggaaga ggttagcgaa cagctcatag aaatcatacc
     4081 gctgtttaat aaggccagat ccttctgcac ccggaagagg tattccactg acaaaattaa
     4141 agtgaatctc aagttcccta cattggcaga cggctgggac cttaataagg agcgggataa
     4201 caaagccgct atattgagaa aagacggcaa gtactatttg gccatcctgg atatgaagaa
     4261 ggatctgtct tcgataagaa cgtcggacga agacgaatcc tccttcgaaa agatggagta
     4321 caaactgctt ccatccccgg ttaagatgct ccctaaaatt ttcgttaagt caaaagccgc
     4381 aaaggagaaa tacggcctta ctgatagaat gctggaatgc tatgataagg gaatgcataa
     4441 aagtgggagc gcattcgatc tcggattctg tcacgagctc attgactatt acaaaaggtg
     4501 catcgcggag taccccggct gggatgtttt tgattttaag ttcagagaaa ctagtgatta
     4561 cggatctatg aaggaattca acgaagatgt ggcaggcgct gggtactata tgtcgctcag
     4621 gaagatcccc tgttccgaag tgtatcggct gttggacgaa aagtcgatat atctgtttca
     4681 aatttacaac aaagattata gtgaaaatgc gcatggtaat aagaatatgc atactatgta
     4741 ttgggaaggg ctcttctctc cacaaaatct tgaatctccg gtctttaagc ttagcggtgg
     4801 ggcagagctt ttcttcagaa aaagcagcat cccaaacgat gcaaaaacgg tgcacccaaa
     4861 ggggagcgtg ctggttccta gaaacgatgt taatgggcgc cggatacctg acagcatata
     4921 cagagagctc acaaggtact ttaaccgggg cgattgcaga atttccgatg aggcgaagtc
     4981 gtacctggat aaagtcaaaa ccaagaaagc cgatcatgac atcgtcaagg accggcgctt
     5041 cacggtcgat aaaatgatgt tccatgtccc aattgcaatg aacttcaaag ctatatccaa
     5101 gccgaatctg aacaagaagg tgatagatgg aattattgac gatcaggacc ttaaaattat
     5161 cgggattgac aggggggaac gcaacttgat atatgtgacg atggtggatc gcaagggcaa
     5221 tatcctttat caagatagcc ttaatatctt gaacggatac gattaccgga aggccttgga
     5281 tgttcgcgag tacgataaca aagaagcaag gaggaactgg actaaggttg aaggaataag
     5341 gaagatgaag gaaggttacc tctcacttgc ggtctccaag cttgcagaca tgattataga
     5401 gaacaacgcc ataattgtta tggaggatct gaatcatggg tttaaggcag gtcgctccaa
     5461 aatcgagaaa caggtttacc aaaagtttga gtccatgctt atcaataagc tcgggtacat
     5521 ggtgctcaaa gacaaatcaa tagatcagtc cggcggagct ctgcacggct accaattggc
     5581 gaaccatgtg actacacttg cctccgttgg taaacaatgt ggcgtcatct tttatattcc
     5641 agctgccttt actagtaaga tagaccccac taccggcttt gccgatctgt ttgctttgag
     5701 caacgtcaag aacgtggcga gtatgagaga gttttttagc aagatgaaaa gcgtcatata
     5761 tgataaagcc gaaggtaaat ttgcgttcac tttcgattat ctggattata acgtgaaatc
     5821 agagtgcggg cggacactgt ggaccgtcta taccgtgggt gagaggttta catactcacg
     5881 ggtcaatagg gaatacgttc gcaaagtccc cactgacata atttacgatg cgcttcaaaa
     5941 ggcaggtatc tccgttgagg gcgacttgcg cgacagaatt gcagagagtg atggggacac
     6001 gctgaaatca attttctatg cgttcaaata tgcgcttgat atgagagtcg agaaccgcga
     6061 agaagattat attcagagtc ctgtgaagaa cgctagtggc gaattttttt gctcaaagaa
     6121 tgcaggtaaa agcttgcctc aggactcgga cgcgaatggg gcttacaata tagcactcaa
     6181 ggggatcttg cagctcagga tgcttagcga gcagtatgac ccgaacgcag aatcgatacg
     6241 gcttccactg atcaccaaca aagcttggct cactttcatg caatcaggaa tgaagacctg
     6301 gaagaacaag cgtcctgctg ccaccaaaaa ggccggacag gctaagaaaa agaagtgaga
     6361 cgactagtgg cggccgccga cgtccgatcg ttcaaacatt tggcaataaa gtttcttaag
     6421 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
     6481 gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag
     6541 agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga
     6601 taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattgatcc cccctcgaca
     6661 gcttccggaa agggcgaatt cgcaactttg tatacaaaag ttgaacgaga aacgtaaaat
     6721 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
     6781 aaaacacaac atatccagtc actatgccat ccagctgata tcccctatag tgagtcgtat
     6841 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     6901 attgcacaag ataaaaatat atcatcatgc ctcctc
//
