LOCUS       sequence-266614-        7047 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 7047)
  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 7047)
  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: pYPQ289 (ErCas12a)
FEATURES             Location/Qualifiers
     source          1..7047
                     /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             6419..6466
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6497..6749
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6794..6917
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6934..6953)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6935..6953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6958..6974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6958..6974)
                     /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 tgaataacgg
     2641 cacaaataac tttcaaaact tcatcgggat ttctagcctg caaaaaacac ttcggaatgc
     2701 actgatcccg acggagacaa cacagcagtt cattgtcaaa aacggtatca ttaaagaaga
     2761 cgagctgcgg ggagagaata gacaaatatt gaaggacata atggatgact attacagggg
     2821 atttatctcc gaaaccctct ctagcataga tgatatagat tggacatccc ttttcgagaa
     2881 aatggaaatt cagctcaaga atggagataa taaggacaca ctgattaagg agcaaacgga
     2941 gtaccggaag gccatccaca aaaagtttgc taacgacgat cggttcaaga atatgtttag
     3001 tgcgaaactt atatctgata tactgcccga gtttgtgatt cacaacaata actattccgc
     3061 atctgaaaaa gaagagaaaa cacaagtgat taaactcttt tcgagattcg ccacctcgtt
     3121 caaagactac tttaaaaaca gagctaactg ctttagtgca gatgatatat cctcgtcgtc
     3181 ttgccacagg attgtcaatg acaatgcgga aatattcttt tcgaacgccc tggtgtacag
     3241 aagaattgtg aaatctttga gcaacgacga cattaacaag atcagtgggg atatgaagga
     3301 ctctttgaag gaaatgtcgc tcgaggagat ttactcgtat gaaaaatatg gtgaattcat
     3361 tacacaagaa ggcatttcat tttataatga tatttgcggc aaagtcaata gctttatgaa
     3421 tctttactgt caaaaaaata aagaaaataa gaacctctat aaattgcaga agctccataa
     3481 gcaaattctc tgcatcgccg atacttctta tgaagtgcct tacaaattcg agtccgatga
     3541 agaggtgtat caatctgtga atggattctt ggataacata tcctcgaagc acattgtcga
     3601 acgcttgcgc aagattggcg acaactataa tgggtacaat ctcgacaaaa tttacatagt
     3661 tagcaaattt tacgaatctg tctcacagaa aacctatcgg gattgggaga ccataaatac
     3721 cgcactcgag attcattata ataatatact ccccggtaac ggaaagtcca aggctgacaa
     3781 agttaaaaag gcagttaaaa acgatctcca aaaatcaata acggagatca atgagttggt
     3841 gagtaattat aagctttgct ccgacgacaa tatcaaggct gaaacgtata tccacgaaat
     3901 tagccatatt ctcaacaact tcgaagcgca ggagttgaaa tacaaccctg agatacacct
     3961 ggttgagagc gagcttaagg ccagcgagct caaaaatgtc ctggatgtga ttatgaatgc
     4021 gttccactgg tgctcagtgt tcatgacaga agaactggtt gacaaagata ataatttcta
     4081 cgcggaactt gaggagattt atgatgagat ctaccctgtt ataagcctgt ataacttggt
     4141 cagaaattat gtcacgcaaa aaccgtactc cacaaaaaag ataaagctga atttcggaat
     4201 ccctactctc gcggatggct ggtccaaatc taaagagtac agtaacaatg ccatcatatt
     4261 gatgagggac aacctttact atttgggaat ctttaatgcg aaaaataaac ctgacaaaaa
     4321 aataatagag ggtaacacct cggagaataa aggcgattat aaaaaaatga tatataatct
     4381 gctgcccggg cccaataaga tgattcccaa ggtttttctt agcagtaaaa ccggggtgga
     4441 gacgtataag ccgtcggcct acattttgga aggctataaa cagaacaagc acatcaaatc
     4501 gagcaaggac tttgatatta ctttctgtca tgacctgata gactatttta aaaactgcat
     4561 agcgattcac ccagaatgga agaacttcgg tttcgacttc tctgacacct ctacatacga
     4621 ggacatatca ggcttttata gagaggtcga gctgcaaggc tacaagatag attggactta
     4681 tatctcagag aaggatatcg acctgctcca agaaaagggc caattgtatc ttttccaaat
     4741 ctacaataaa gatttttcta aaaagtccac agggaacgat aatctccaca cgatgtacct
     4801 taaaaacctc ttttcggaag aaaacctgaa ggacattgtg cttaagctga atggggaggc
     4861 cgagatattc tttaggaaga gtagcataaa gaacccgata atccataaaa aaggttccat
     4921 actggttaat cggacctatg aggcagaaga aaaagaccag ttcggaaata tccagattgt
     4981 cagaaaaaat ataccagaga atatctacca ggaattgtac aaatatttca acgataaatc
     5041 agataaggaa ctctcggacg aagcagcgaa attgaagaac gtcgttggac accacgaggc
     5101 cgccactaat atcgttaaag attacaggta cacgtacgat aagtattttc tccacatgcc
     5161 aatcactata aactttaaag ctaataagac cggctttatt aacgatcgca tccttcagta
     5221 tatcgctaag gagaaggacc tgcacgttat aggaatagac cgcggcgagc gcaatcttat
     5281 ctacgtgagc gtgatcgata cctgtgggaa tatagtcgag cagaagtctt ttaatatcgt
     5341 gaacggctac gactatcaga taaagctgaa gcagcaagag ggcgctagac agatcgccag
     5401 gaaggaatgg aaggagatcg gtaaaataaa agaaattaag gagggttacc tcagtttggt
     5461 tatacatgag atatcaaaaa tggttattaa gtataatgcg attattgcga tggaagacct
     5521 gtcatatggg tttaaaaaag ggcggttcaa ggttgagcgc caggtctatc aaaagtttga
     5581 gactatgctg attaataagc tcaattatct tgttttcaaa gacatcagta tcacggaaaa
     5641 cggaggactt ctcaaggggt accagttgac ttatatccca gacaagctta aaaatgttgg
     5701 tcaccagtgt ggatgtatat tttatgtgcc tgcggcctac acatctaaga tcgacccaac
     5761 aactggcttt gttaatattt ttaaattcaa agaccttacg gtggatgcga aaagagagtt
     5821 cataaaaaaa ttcgactcga tcagatatga ctctgagaag aatctcttct gttttacttt
     5881 cgactataat aatttcataa cacaaaatac cgtcatgtcg aaatcaagct ggtcagtcta
     5941 cacatatggc gttcgcataa agcgccgctt cgttaatggc aggttctcga atgagtccga
     6001 tacaatcgac attacgaagg atatggagaa aacactcgag atgacggata taaattggag
     6061 ggatgggcat gatctgagac aggacataat agactatgag atagttcagc acatattcga
     6121 gatcttcaga ctcactgttc agatgcggaa ctccctcagt gaacttgaag acagggatta
     6181 tgataggctt atttctcctg tcctgaacga aaacaatatt ttctacgact ctgctaaagc
     6241 aggggatgct cttcctaagg atgcagacgc aaatggcgca tactgcatag ctcttaaagg
     6301 gctctatgag atcaagcaga ttacagaaaa ctggaaagaa gacgggaagt tttccagaga
     6361 caaattgaag atctccaaca aggactggtt cgattttata caaaataaaa ggtaccttaa
     6421 gcgtcctgct gccaccaaaa aggccggaca ggctaagaaa aagaagtgag acgactagtg
     6481 gcggccgccg acgtccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     6541 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     6601 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     6661 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     6721 gcgcgcggtg tcatctatgt tactagatcg ggaattgatc ccccctcgac agcttccgga
     6781 aagggcgaat tcgcaacttt gtatacaaaa gttgaacgag aaacgtaaaa tgatataaat
     6841 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     6901 catatccagt cactatgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     6961 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     7021 gataaaaata tatcatcatg cctcctc
//
