LOCUS       sequence-272072-        7008 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 7008)
  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 7008)
  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: pYPQ283 (MlCas12a)
FEATURES             Location/Qualifiers
     source          1..7008
                     /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             6380..6427
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6458..6710
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6755..6878
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6895..6914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6896..6914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6919..6935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6919..6935)
                     /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 cctgcggctc tgtttcaaga
     2641 tttcacacac ctttacccac tgtctaaaac ggtgcggttt gagcttaaac ccatcggaaa
     2701 aacgctggaa catattcatg cgaagaactt tttgtcgcag gatgagacaa tggcagacat
     2761 gtaccaaaaa gtgaaagcca tactggacga ttaccataga gacttcatta cgaagatgat
     2821 gagtgaggtg actcttacaa agctcccgga gttttacgag gtttacctcg ctctgaggaa
     2881 gaatccgaaa gacgataccc tccagaaaca acttaccgaa attcagaccg cgctccggga
     2941 ggaagttgtg aagcccatag attccggagg taaatacaag gctggttatg agcgcttgtt
     3001 tggggcgaaa ctgtttaaag acggaaaaga gttgggagat ttggcgaaat ttgtcatcgc
     3061 ccaggaaggg gaatcgtctc cgaaacttcc acagatcgcc cactttgaaa agttttcaac
     3121 atacttcacg gggttccacg ataaccgcaa gaacatgtac tcgtctgacg ataaacacac
     3181 agctattgcc tacaggctta tccatgaaaa tttgccccgc tttattgata acttgcaaat
     3241 tttggttact attaagcaga agcactcagt cctttacgat cagattgtga acgagctcaa
     3301 cgccaatggc ctggacgtgt ctctcgcctc gcatctcgac ggctaccata aacttttgac
     3361 tcaagaaggc attaccgcgt acaataggat tattggcgaa gtgaacagtt atacaaacaa
     3421 gcataatcag atatgtcaca agtctgaaag aattgcaaag ctgagaccgc ttcataagca
     3481 aatacttagt gatgggatgg gagtgagttt cctcccatct aaatttgcag atgactcgga
     3541 aatgtgtcaa gcggtgaatg aattctacag acactatgcc cacgtgttcg ccaaagtcca
     3601 aagtctcttt gaccggtttg acgattacca aaaagacggc atttatgttg aacacaagaa
     3661 tctcaacgaa ttgagcaagc aggcctttgg agatttcgca ctccttggca gggtccttga
     3721 cggctactat gtcgacgttg tcaatcctga gttcaatgac aaattcgcta aggctaagac
     3781 agataacgcc aaagagaaac ttaccaaaga aaaagacaaa ttcattaagg gagtgcattc
     3841 attggcttct ctcgagcagg caatcgaaca ctatatagcg gggcatgatg acgaaagcgt
     3901 tcaggcggga aaacttggcc aatacttcaa gcacggcctc gctggagttg ataatcctat
     3961 acaaaagatt cataatagtc attctacgat aaagggattc cttgagaggg agaggccagc
     4021 cggagaacgc accctcccta aaataaaatc agataagtct ctggagatga ctcaactcag
     4081 acagctgaag gagctgctgg acaacgccct caatgtcgtc cacttcgcaa agctgcttac
     4141 aaccaaaacg accctggata accaagatgg aaatttctac ggagaattcg gggcccttta
     4201 tgatgagttg gctaaaatcg caaccttgta caacaaggtg cgggattatc ttagccaaaa
     4261 accgttcagt acagaaaaat acaaacttaa ctttgggaat ccgactctct tgaatggatg
     4321 ggacctcaat aaggagaagg ataattttgg cgtcatcctg cagaaggacg gctgctacta
     4381 tctcgcactg cttgataaag ctcacaaaaa agtctttgat aatgcgccca acacgggtaa
     4441 gtcggtgtat cagaaaatgg tgtataaact tttgcctgga tcaaataaga tgctcccaaa
     4501 agtttttttt gcgaaatcga accttgatta ctataatcct agtgccgaac tcctcgacaa
     4561 gtatgcccaa ggcactcaca agaagggaga caacttcaac ctgaaagatt gccatgcatt
     4621 gatcgacttt ttcaaagcgt ccattaacaa gcaccccgaa tggcaacatt tcggttttga
     4681 atttagcttg acgagctctt accaggacct gtcagacttc tacagggagg tggagccgca
     4741 aggctatcaa gttaagttcg tcgacattga tgctgactac atcgacgaac ttgtggaaca
     4801 aggtcaactc tacctgtttc aaatatataa caaggatttt agtcctaagg cgcacggaaa
     4861 acccaacctt catacgctct acttcaaagc tttgttctca gaagataacc tggctaaccc
     4921 aatatataag ctcaacggag aagccgaaat cttctaccgg aaagcttcgc tcgatatgaa
     4981 cgagacaacg atccatcgcg cgggtgaggt gcttgaaaat aagaacccgg ataacccgaa
     5041 agagaggcag tttgtgtacg atataatcaa agacaaacgc tacactcagg ataagtttat
     5101 gctgcatgtt ccaattacaa tgaactttgg ggttcaagga atgacgatca aggagtttaa
     5161 caaaaaggtt aaccagtcca tacaacagta tgacgaagtt aatgtcattg gaatagacag
     5221 aggcgagcgc catctcctgt accttacagt tattaattct aagggagaaa tactcgagca
     5281 acggtcgctc aacgacatca taactaccag cgcgaacggc acccagatga caacaccata
     5341 tcacaagatc ctggacaaga gggagatcga gcggctgaat gctagggtgg gttggggaga
     5401 aatagagaca atcaaggagt tgaagtctgg ttatctttcg catgtcgtgc accaaatatc
     5461 gcagctgatg cttaaataca atgctatagt tgtccttgaa gatttgaatt ttggcttcaa
     5521 acggggtcgc ttcaaagttg aaaaacagat atatcagaat tttgaaaacg ctcttattaa
     5581 gaaactgaat catctggttt tgaaggataa agcggacaat gaaatcggct cttacaaaaa
     5641 tgcgcttcag ttgactaaca attttaccga tctcaagtct atagggaagc agacaggttt
     5701 cctcttttac gtgcctgctt ggaacacgtc aaaaattgat cccgttaccg gttttgttga
     5761 cctgttgaaa ccgcggtacg aaaatattgc acaaagtcaa gcattttttg ataagttcga
     5821 caagatttgc tataatgctg acaaaggcta cttcgagttt catatagact atgcaaagtt
     5881 cacggacaag gcaaaaaact cgcgccagat atggacgatt tgctcgcatg gagacaagcg
     5941 ctatgtgtat gacaagacag ccaatcagaa caagggagca actattggga tcaatgttaa
     6001 tgatgagctc aagtcgttgt ttgcccgcta ccggattaac gacaagcagc caaacctggt
     6061 catggatatc tgtcaaaata atgataaaga gtttcacaag tcgctcacgt accttcttaa
     6121 agctctcttg gcattgcggt acagcaatgc cagttccgac gaagatttta tcctgagccc
     6181 ggtggccaac gataaggggg tgtttttcaa ttctgcactg gctgatgata cccagccgca
     6241 gaatgcggac gcaaacggag catatcatat cgcgttgaag ggactttggc ttctcaatga
     6301 gcttaagaac tcggatgacc ttgataaagt gaaactcgcc atcgacaatc agacatggct
     6361 taatttcgca cagaacagaa agcgtcctgc tgccaccaaa aaggccggac aggctaagaa
     6421 aaagaagtga gacgactagt ggcggccgcc gacgtccgat cgttcaaaca tttggcaata
     6481 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     6541 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     6601 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     6661 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattgat
     6721 cccccctcga cagcttccgg aaagggcgaa ttcgcaactt tgtatacaaa agttgaacga
     6781 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     6841 acataatact gtaaaacaca acatatccag tcactatgcc atccagctga tatcccctat
     6901 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     6961 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctc
//
