LOCUS       sequence-266608-        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: pYPQ285 (Lb5Cas12a)
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 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 cctgcggctt actatgaatc
     2641 cctgaccaaa caataccctg tttccaaaac aatccgcaat gagctgattc ctataggtaa
     2701 aaccctggat aacatccgcc aaaataacat cttggaatca gacgttaaaa gaaagcagaa
     2761 ttacgaacat gtcaaaggta tacttgatga ataccacaaa cagctcataa acgaagcgtt
     2821 ggataactgt acgcttccct cactgaaaat agcggctgag atctacctca aaaaccagaa
     2881 ggaggtgtca gacagagaag atttcaacaa aacccaggat ctgttgcgca aggaggttgt
     2941 ggagaaactc aaggcgcatg aaaattttac taagatagga aaaaaagata ttcttgattt
     3001 gttggagaag cttcctagca tatccgagga cgactacaat gccctggaaa gtttcagaaa
     3061 cttttacaca tacttcacat cgtataataa ggtccgggaa aatctgtata gcgataaaga
     3121 gaaaagttct actgttgcgt acaggcttat caatgagaat tttccaaagt ttctcgacaa
     3181 cgtcaaatca tatcggttcg tcaaaactgc gggcattttg gctgacggat tgggagagga
     3241 ggagcaggac agcctgttca tagtggagac tttcaacaag acattgaccc aggatggcat
     3301 tgatacatat aactcccagg tcggcaagat aaactcctcg ataaacctct acaaccagaa
     3361 aaaccagaag gcaaacggct tccggaaaat cccaaagatg aaaatgcttt ataagcagat
     3421 cttgagtgat cgggaggagt ctttcatcga tgaatttcag tcagacgaag ttcttatcga
     3481 caacgttgag agttacggct ctgtgcttat tgagagcctc aagtcgtcaa aggtttctgc
     3541 attttttgat gcacttcggg agagtaaagg taaaaacgtt tacgttaaga atgacctggc
     3601 gaaaacagca atgtcaaaca tagtttttga gaactggagg accttcgatg accttctgaa
     3661 tcaagagtac gatttggcga acgaaaataa aaagaaggac gacaagtact ttgagaagag
     3721 gcaaaaggag ctgaaaaaga ataaatcgta ttcgttggaa catctttgca acctctctga
     3781 ggattcctgc aacctgatag aaaactacat ccaccagatc agtgacgata ttgaaaacat
     3841 tattattaac aatgaaacct ttttgcggat agtgattaat gagcacgatc gcagtagaaa
     3901 acttgctaaa aatagaaaag ctgttaaagc aataaaggat ttccttgaca gtattaaggt
     3961 gctcgagcgg gagttgaaac tgatcaattc ttcaggacaa gagttggaaa aagaccttat
     4021 cgtctatagc gctcacgagg aacttctggt ggaactgaaa caagttgatt cgctttataa
     4081 catgacgagg aactacctga ccaaaaagcc cttttctact gagaaagtca aactcaattt
     4141 taatcggtcg acgcttctga atggctggga ccgcaacaaa gagactgata acctcggggt
     4201 tctcttgctg aaagatggca agtattatct gggtataatg aatacatcag caaataaggc
     4261 attcgttaac ccgcctgtgg ctaaaaccga gaaggttttt aaaaaagtgg actacaagct
     4321 gttgccagtg ccgaaccaga tgttgcccaa ggtttttttt gcgaaaagca atattgattt
     4381 ctataatcca tcgagcgaga tatattccaa ctacaaaaaa ggtacccata aaaagggtaa
     4441 catgttttca cttgaggact gccacaacct gattgatttc tttaaagaga gtattagtaa
     4501 gcacgaagat tggagtaagt ttggattcaa attttctgat actgcgagtt acaatgatat
     4561 aagtgaattc tatcgggaag tcgaaaaaca gggttacaag cttacgtaca ctgacataga
     4621 tgaaacatac attaatgacc tgattgagcg caatgagttg tacctgtttc agatctataa
     4681 caaagatttt tccatgtata gcaaaggcaa gctcaacttg cataccctgt attttatgat
     4741 gctcttcgat caaaggaata tcgatgacgt tgtttataag ctgaatggtg aagcagaagt
     4801 tttttaccgc ccagcctcga tcagtgaaga tgagctgatc attcacaaag cgggtgaaga
     4861 gattaagaat aagaacccta atcgggccag gacgaaagaa acttcaacgt tttcctatga
     4921 tattgtcaaa gacaaaaggt actcgaagga taagtttacc cttcacattc ccataaccat
     4981 gaattttggt gtcgacgaag tcaagcggtt taacgacgcc gtgaattccg cgatccggat
     5041 agacgagaat gtgaatgtca ttgggattga cagaggtgaa cgcaatcttc tttacgtggt
     5101 ggtgatagac tctaaaggga atatattgga gcaaatttct ctgaactcaa tcattaataa
     5161 agaatacgat attgaaacag actatcatgc ccttctcgac gagcgggagg gcgggaggga
     5221 taaagcacgg aaggactgga acacagttga gaatatccgg gacctcaaag ccggatacct
     5281 ttcccaggtg gtcaatgttg tcgcaaaatt ggtcttgaag tacaatgcca ttatatgcct
     5341 tgaagatttg aactttgggt tcaaacgcgg taggcaaaaa gttgaaaaac aagtttacca
     5401 gaaattcgaa aagatgctta tagataagct caactacctg gttattgaca agagtagaga
     5461 gcagacgtcg cccaaagaac tcgggggggc gctgaatgcg ttgcaactga cctcgaagtt
     5521 caaatcgttc aaagaattgg ggaaacagtc cggcgttatc tattacgtgc ctgcttatct
     5581 cacgtctaag atagatccta ccacaggctt cgcgaatctt ttttatatga agtgtgagaa
     5641 tgtcgagaag tctaagcggt ttttcgatgg atttgacttc atcaggttca atgcgctcga
     5701 aaatgttttt gagtttggat tcgactaccg gagctttaca cagagagcgt gcggtataaa
     5761 cagtaaatgg acagtctgca caaatgggga gagaattata aagtatagga accccgataa
     5821 aaataacatg ttcgacgaaa aggtggtggt tgttactgat gaaatgaaga acctgtttga
     5881 acagtacaaa ataccttatg aggatggaag gaacgtgaag gacatgatta tttcaaacga
     5941 ggaggcggag ttctaccgca gattgtaccg cctgcttcag cagaccctcc aaatgcggaa
     6001 ttcgacctca gatggaacga gagattatat tatatcacca gtgaaaaaca agcgcgaggc
     6061 gtatttcaac tctgagcttt ccgatggttc ggttccgaaa gacgcagatg caaacggagc
     6121 atacaacatc gcaaggaaag ggttgtgggt gttggaacaa atccgccaga aatccgaagg
     6181 cgaaaaaatt aacttggcta tgaccaatgc tgagtggctt gaatacgcgc agacgcacct
     6241 tctgaagcgt 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
//
