LOCUS       sequence-266602-        7149 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 7149)
  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 7149)
  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: pYPQ282 (TsCas12a)
FEATURES             Location/Qualifiers
     source          1..7149
                     /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             6521..6568
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6599..6851
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6896..7019
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7036..7055)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7037..7055)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7060..7076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7060..7076)
                     /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 cctgcggcta ccaaaacctt
     2641 cgattcagaa ttctttaatc tttactcact ccagaaaacc gtccgcttcg aattgaaacc
     2701 cgttggcgag actgcatcat tcgtcgaaga ttttaagaat gagggattga agcgggtcgt
     2761 gagcgaagac gagcggagag cagtggacta tcagaaggtg aaggagatta ttgatgatta
     2821 ccatcgcgac tttattgagg agagccttaa ctacttccca gagcaggtgt ccaaagacgc
     2881 acttgaacag gcatttcacc tctaccaaaa gctgaaggca gcgaaggtgg aggagcggga
     2941 aaaagctctg aaggaatggg aagctttgca gaagaaattg cgggagaaag ttgtcaagtg
     3001 cttctcagat tcgaataaag cgagatttag cagaattgac aaaaaggagc ttatcaagga
     3061 ggatcttatt aactggcttg tcgcccagaa tcgggaagac gatattccga ctgtcgagac
     3121 gtttaacaat tttacgactt acttcacggg ctttcacgag aataggaaga atatttactc
     3181 caaggacgat cacgctactg cgattagttt tcgcctcatc catgaaaatt tgcccaaatt
     3241 tttcgacaat gttatctcct ttaataagct caaggaagga ttcccggaac tgaaatttga
     3301 taaagttaag gaagatttgg aggtcgatta tgacctcaaa cacgcattcg agatagaata
     3361 tttcgtcaat tttgttacgc aggccgggat agaccagtac aactacctgc ttggcggtaa
     3421 gacgctcgaa gacggaacca aaaagcaggg tatgaatgaa cagatcaacc tgtttaaaca
     3481 gcagcagacc cgcgacaagg cacggcagat tcccaagttg ataccgctct ttaagcagat
     3541 tttgagcgaa cggacagaat cgcaatcgtt catcccgaag caatttgagt ccgatcaaga
     3601 gctcttcgac tcattgcaga aattgcacaa caactgccag gacaagttca cagtccttca
     3661 acaggccata ttgggattgg ccgaggccga tctcaaaaag gttttcataa agacatcgga
     3721 cttgaatgct ctttcgaata cgatattcgg aaattattcc gtgttttccg acgcgctcaa
     3781 cctctataaa gagagtctta aaacgaaaaa agcccaagaa gcgtttgaaa agctgccggc
     3841 tcacagcata cacgatctga tacagtatct ggaacaattc aattccagcc tcgacgctga
     3901 gaagcagcaa tcaacggaca ccgttctgaa ctattttatc aagaccgatg aactgtattc
     3961 gagattcata aaaagtacat cagaagcctt cacgcaggtc cagccattgt tcgagttgga
     4021 agccttgtcg tctaaaagac ggccccctga gtctgaagat gagggcgcaa aaggccaaga
     4081 gggattcgaa caaataaagc gcatcaaggc ctatctggat actcttatgg aagcggttca
     4141 ctttgctaag cccctctatc ttgtcaaagg gaggaagatg attgagggtc tggataaaga
     4201 ccaatcattt tacgaagcat tcgaaatggc gtaccaggag cttgaaagtt tgattatacc
     4261 gatatataac aaagcaaggt cgtacctttc aaggaaacct tttaaggcag ataaattcaa
     4321 gataaatttc gataacaaca ctctcttgag cggttgggat gctaataaag agaccgctaa
     4381 cgccagtatc ttgtttaaga aagatggcct ttactacctg ggtatcatgc ctaaaggaaa
     4441 aacgtttctc ttcgattatt ttgtgagttc tgaggattct gagaaattga aacagcggag
     4501 acaaaagact gcggaggaag ctctcgcgca agacggtgag tcgtattttg aaaaaatcag
     4561 gtacaagttg ttgccgggag ctagcaagat gctgccaaaa gttttcttct cgaataagaa
     4621 tataggtttt tacaacccta gcgatgacat cctccggatc aggaacacag cgtcccacac
     4681 aaagaacggt accccacaaa agggacattc caaagtggaa ttcaacctga acgattgtca
     4741 taagatgata gatttcttta agtcttctat acagaaacat cccgagtggg gcagttttgg
     4801 gtttactttc agcgacacta gtgatttcga agatatgtct gctttctaca gggaggttga
     4861 aaatcaaggc tatgtgatct cttttgataa aattaaagaa acatatattc aatcgcaagt
     4921 cgaacaaggg aacctctacc tcttccaaat ctacaataag gactttagtc cttattccaa
     4981 ggggaaaccc aaccttcaca cactctattg gaaggcactt ttcgaagagg cgaatcttaa
     5041 taatgtcgtg gcaaaactga acggcgaagc agaaattttt ttcagaagac attccataaa
     5101 agcaagcgac aaagttgtgc accctgctaa tcaagcaata gacaacaaga atccgcatac
     5161 cgaaaagaca caaagtacct tcgagtacga tctggtgaaa gataaaagat acacacagga
     5221 caagtttttt ttccacgttc caatctcctt gaactttaag gctcaaggtg tttcgaagtt
     5281 taacgacaag gttaacgggt ttctcaaagg gaacccggac gtgaacatca ttggcattga
     5341 tcggggcgag cggcatttgt tgtactttac tgttgtgaac cagaagggtg agatattggt
     5401 tcaggagtca cttaatactc tgatgtcaga taaaggacac gtgaatgact accaacagaa
     5461 gcttgataag aaagaacaag aacgggatgc cgctaggaag tcatggacaa cagtcgaaaa
     5521 tattaaagaa ttgaaggagg gttacctgag ccatgttgtt cataagctgg cacatctcat
     5581 aatcaaatac aacgcgatcg tctgtctgga agatcttaat ttcggtttca agaggggtcg
     5641 ctttaaagtg gaaaagcaag tctaccagaa attcgaaaag gcccttattg acaagctgaa
     5701 ctatctcgtt tttaaagaga aggaattggg ggaggtcggg cattatctga ctgcatacca
     5761 actgactgct cctttcgaat ccttcaaaaa gttgggtaag cagtctggta tactctttta
     5821 tgtgccggct gattacacct ccaagatcga ccccaccaca ggatttgtta acttcttgga
     5881 tctgagatat caaagcgtcg agaaggctaa gcaacttttg tccgatttca acgctattcg
     5941 ctttaattct gtccagaatt atttcgagtt tgaaattgac tacaaaaagc tgactcctaa
     6001 acggaaagtt ggcacgcaga gcaagtgggt catttgtact tatggtgatg tcaggtacca
     6061 aaatcgcaga aaccagaaag gccattggga gaccgaagag gtcaacgtta cggagaaact
     6121 caaagcgctc tttgccagtg attcgaagac cactaccgtc atcgactatg ccaatgatga
     6181 taaccttata gatgtcatac ttgagcagga taaagcgagc ttctttaaag aattgttgtg
     6241 gctcctgaag ctgacaatga ccttgaggca tagtaaaatt aagagtgaag atgactttat
     6301 tctgtctcca gtcaagaatg aacagggcga gttttacgac tccaggaaag cgggggaagt
     6361 ctggcctaaa gatgccgacg ctaatggcgc atatcatatt gctctgaaag gtctttggaa
     6421 tctccaacaa ataaatcaat gggaaaaagg caaaaccttg aacttggcta ttaaaaatca
     6481 agattggttc tccttcatcc aagaaaaacc ctatcaagag aagcgtcctg ctgccaccaa
     6541 aaaggccgga caggctaaga aaaagaagtg agacgactag tggcggccgc cgacgtccga
     6601 tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat
     6661 gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat
     6721 gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc
     6781 gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat
     6841 gttactagat cgggaattga tcccccctcg acagcttccg gaaagggcga attcgcaact
     6901 ttgtatacaa aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     6961 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgc
     7021 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     7081 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     7141 tgcctcctc
//
