LOCUS       sequence-176291-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AsCpf1 Gateway entry plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  AUTHORS   Tang X, Lowder LG, Zhang T, Malzahn AA, Zheng X, Voytas DF, Zhong Z,
            Chen Y, Ren Q, Li Q, Kirkland ER, Zhang Y, Qi Y
  TITLE     A CRISPR-Cpf1 system for efficient genome editing and 
            transcriptional repression in plants.
  JOURNAL   Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18.
  PUBMED    28211909
REFERENCE   2  (bases 1 to 7214)
  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 
            Plants."; date: "2017-02-17"; volume: "3:17018. doi"; pages: " 
            10.1038/nplants.2017.18"
COMMENT     Sequence Label: pYPQ220 (AsCpf1)
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..66)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(49..65)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(58..80)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      129..215
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      307..334
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(493..510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(664..1252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(744..763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1345..2136)
                     /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"
     primer_bind     2567..2583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2567..2583
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2588..2607
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2588..2606
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(2624..2747)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      complement(2792..3044)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(3113..3160)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3161..7078)
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     CDS             complement(7103..7123)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(join(7148..7214,1..33))
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggtcactatc agtcaaaata aaatcattat ttggggcccg agcttaagac tggccgtcgt
       61 tttacaacgt cgtgactggg aaaacatcca tgcaagcgtt aacgcgagag tagggaactg
      121 ccaggcatca aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt
      181 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
      241 tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
      301 ctaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctggct
      361 cgcggtacgc gtattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
      481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1261 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
     1321 ttttggtcat gggtggctcg agggttattt gccgactacc ttggtgatct cgcctttcac
     1381 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     1441 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     1501 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     1561 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     1621 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     1681 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     1741 agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg
     1801 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     1861 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     1921 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     1981 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     2041 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     2101 ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg
     2161 actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     2221 acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca
     2281 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     2341 gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca
     2401 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag
     2461 cgaagtcgag gcatttctgt cctggctggt ccagaggagg catgatgata tatttttatc
     2521 ttgtgcaatg taacatcaga gattttgaga cacgggccag agctgccagg aaacagctat
     2581 gaccatgtaa tacgactcac tataggggat atcagctgga tggcatagtg actggatatg
     2641 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     2701 atttatatca ttttacgttt ctcgttcaac ttttgtatac aaagttgcga attcgccctt
     2761 tccggaagct gtcgaggggg gatcaattcc cgatctagta acatagatga caccgcgcgc
     2821 gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta aatgtataat
     2881 tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta catgttaatt
     2941 attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag accggcaaca
     3001 ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatcggacgt caccggtgtt
     3061 taaacgctag ctctagatca ggtggatccg gcggccgcca ctagtcgtct cacttctttt
     3121 tcttagcctg tccggccttt ttggtggcag caggacgctt attgcggagc tcttggatat
     3181 aggcaagcca atcctggttg ctgatgccgt tctgcagctt aaggtccttg ctctccttga
     3241 gatggttcaa caacagttgt cctttcaatg ctatgtggta tgcgccgtta gcatcggcat
     3301 ccatcggcca ctcaggattc tgaaatctag aatcgaagca cacgccattg agatcacgca
     3361 caggggagtt gatgtaatcc tccccagtgg cagcatttga atttctcatt tgcagcacgg
     3421 atctgatgag cgctaccatc gtgtcaatag cgtggctatc atcgttctcg agcagtttgg
     3481 ggaggatatt tgacccgtcg cggaaaacga tgcctttctc ctccaggagg gcgatgagtt
     3541 cgtttgcggg gtacaggtca cggtaacgcc cggtgaatct atggttctca attacaggga
     3601 caatcctctt gcccgcgatg aaaggcgtgc ctttggcgtc gaattgggtt tcatttttct
     3661 caaaaacgat atcccaagcg ggcatgaagc cagggagtcc ccgctggaag gagaggttcc
     3721 gattcatctt gaaatgcagg atgaagtctc cggtttttac atcatagtgc aggaagtcga
     3781 aaccctccag gaaatgctta cgcgactcat ggttcttgat ggtcttccag acaaacgggt
     3841 ccacgaatcc ggtcaaggga tcaatctttg aggtatatgg ggccgggaca tagaaaagga
     3901 agccgctctg ggtgcccatc ttcgcaaatg acgtaaactg gtcggtcagc tgatacggat
     3961 tgaggacccc cccgaccttc tccgccggat agtccttgag caccaggcaa ttgagcttgt
     4021 cgataagcat tttctcgaac tgttggtaca ccgccttctc ggcgatgcct gtgcgcttgg
     4081 atttgaatcc gaaattgaga ttctcgagga ccacgacggc ttggtagtgg atcatgagat
     4141 ccactatctc atggatgacc tggctgagat agccctgctt caggtctttg atcgtgccca
     4201 ctacggacca ggcctgcctc gcggcgacgc gttccttctc ccggttgtcg agcttcttct
     4261 ggtagtcgaa ctgctggatg gtattgagcg accgttgctc gagaatcttt ccggtggagt
     4321 cgatgacggt gatgtagatg aggttccgct ccccccggtc aatcccgatt atgggtgtct
     4381 ctgggtgctc cttgagatac gcgttgaccc gctggttgaa cttggacggg ctattcgcgg
     4441 cctggtagtt cagcgtgatg gggacatgga aaaagaactt gtcagaggta aaccgccgat
     4501 cctttatgat ctcgtggctg acctccttcg tgatgacgtt tggcaggagc gcgcgggcct
     4561 cgtcgctgag gtcgtgtgag aggcggtggt tcacgtaatc gtacagctcc tgatacagtg
     4621 tatcagggat tggcgtcttt tgatctttca gcttcttgtt cagcatcttc tcaccgaggc
     4681 gatgagccat gcgcttcatc ctggacttcg gcctgtagaa gagttctgcc tggccgttca
     4741 gctttatgct cgtcttcgcg agattttccg ggctgaagag gcctgtccag tagagggtgt
     4801 gcaggtttgg cttcccgtgg tggcccttcg cgaagtcctt gttatagatt tggaaaagat
     4861 acaacttgcc ggtttcgacg gcgtccatga tctccttttc cgcgatacgc tgaaacgaga
     4921 tgtgatacag gagtgggttg agttcggcgt agtactcccc gaggtccttg tactgggaag
     4981 aggggcggag ggaagagaga tcgatggagg ttgtcttggt gtacttgctg aggaaatcgc
     5041 gcgtgaagtc gatccacttg cagagcgcct ctctataccc cttttgatcg cccgtcttct
     5101 tggcgtaggc ggtctggaac ttcttgggtt ccttttctgg attattcaga tcgtatatct
     5161 ccttggtgat ctcgagcggc tcgatgaagt tgttggagag gaggatgggg gtggtatggg
     5221 tctgaaagtg cgccgtcacc gccttaagct gagtgctgca cttcggtatc atctttgcag
     5281 cgtcaggaaa ataatcgtag tacatcttgt cgaagccctc ggaggtcttc tcagttggct
     5341 cgaagctgag cgccttgtac ctgccttttt gtttcggcat aattcccaaa taatacaacc
     5401 catttttcac gaaaagaatg gctccgttgt tcttctcttt gttaacgtcc cacccgctgg
     5461 ccagggtggg catttggaag ttcagcttga acttctccac ggaatacggc ttcttggtcg
     5521 cgtaattgcg cgccttatta tagaagctca agctaggttc catctcaagc ttgatcccgg
     5581 tgaggcgggc gctaaactcc ggatcgacct cgttggactc gtccacggcg aaccagtcga
     5641 gaaggtggta cagccccagg aggctgtcca gctgggactt gaggatttcc ttttcctctt
     5701 gctttttcag ggtcgtcggc agcggctggt ccagggccgc gtgggcgtga gagaggatct
     5761 ccgaggtctt ctgcttgaaa gcctcggaca gctccttgcc cgcggcggag atgatctcct
     5821 gcaggttaat gtcctcgtgc ttgaggctcc tttgcacttt ctctttcgcc gacttcgtga
     5881 tcttgccggt gagctcgctg atgcgccgct cgtagagggc gttgcggagg gtgtcccagt
     5941 ggtcgcacag ggcgctggag atggtttcca gcttcttatg ggagatgaag atgtgggtga
     6001 gatcgatgct attgagctca ttgaacagcg cctcggcggt ttcgagcaca ttctcgttgc
     6061 ggaggagggt cttgtacttg cagaaggact ggatgacctc ctcgtcgctc ttgaactcct
     6121 cgaggatgaa ggacaacgtg ttgcggtcgg agaggatctg cttaaacagg gggataaacc
     6181 tatgtgggag tgaggcaatg atgtgcgcgg tttcgtcgtt cttctggatg gcgagattga
     6241 gcacctcgtt caggcccttg atcttctcgg tgccggcctc gcgggagatg ccgccgagga
     6301 gctggttgta gaggtcgatc tgggtctgcg tcagcagctg attgtagaag gggaaggaga
     6361 acacctcttc gatcgaggtg gacacgaaga tcccgatggc cttcttcacg ttctcgaagt
     6421 gctcgcggag ggacggcacg gccgtgatga ggcgggtgaa gatgtggcag ttctccttga
     6481 acttcgggaa gttgtcttgc acgatcctat gcgggatggc ggtgctaatg tcctcggcgc
     6541 tgaacacgtt tttgcggttc tcgtaaaagc cgctaaagta tgtcgtaaac ttgtcgaacg
     6601 accggaggag ggcgttctca tgctctgtgg tcgtgacggt gcccagttgc ttcagcactt
     6661 tgccattgaa cagctcggcc ttaaacagcc ccttatagat ttcggcgtgg cgcttgttga
     6721 tggcgtcggt gagattatct gtgcgcccga tgaagtagtc gtggatagcg ttgcgatacg
     6781 tagcctgttc ttcgatgagc gcgttgcgtg tctcctccgt cttctccttg cggtaggagt
     6841 cgatggcggc cgagaggttc tcccagtcga gctgcacgag ctgcaggcac tggtccgcgt
     6901 aggtcttgta gatccggtcg atgatcggct tgagctcctt gtagtggtcg ttccgcgctt
     6961 tgtcctcctc gataaagccc tgctcctgga tgtgcttgag ggtcttgccc tgcggaatca
     7021 gctcgaaccg cagcgtcttg ctgacctggt agaggttggt gaacccctcg aactgcgtag
     7081 ccgcaggcac cccgtgaata ccaaccttcc gcttcttctt aggagccatg gtgaagggcg
     7141 aattcggagc ctgctttttt gtacaaagtt ggcattataa aaaagcattg ctcatcaatt
     7201 tgttgcaacg aaca
//
