LOCUS       sequence-78695-e        4033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with attL1 and attR5 sites flanking Zif268:FokI
            coding sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4033)
  AUTHORS   Baltes NJ, Gil-Humanes J, Cermak T, Atkins PA, Voytas DF
  TITLE     DNA Replicons for Plant Genome Engineering.
  JOURNAL   Plant Cell. 2014 Jan;26(1):151-63. doi: 10.1105/tpc.113.119792
  PUBMED    24443519
REFERENCE   2  (bases 1 to 4033)
  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"; doi: "10.1105/tpc.113"; 
            journalName: "Plant Cell"; date: "2014-01"; volume: "26"; issue: 
            "1"; pages: "151-63"
COMMENT     Sequence Label: pNJB99
FEATURES             Location/Qualifiers
     source          1..4033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             16..36
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             307..891
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="LVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVM
                     EFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRY
                     VEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLS
                     VEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     terminator      905..1157
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    1202..1325
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1342..1361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1343..1361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1366..1382)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1366..1382)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1813..2604
                     /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      2697..3285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3186..3205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3439..3456
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3615..3642)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3734..3820)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3869..3891
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3883..3900
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3884..3900
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3916..4015
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atggcttcct cccctccaaa gaaaaagaga aaggttcttg aattgcctta cgcttgtcca
       61 gttgagtctt gtgatagacg ttttagtagg tcagacgaac tcactaggca tattagaatc
      121 cacacaggac agaagccttt ccagtgtaga atctgtatga ggaactttag tcgtagcgat
      181 catttgacta cacatatcag aacccatact ggtgagaaac cattcgcatg tgacatctgt
      241 ggtcgtaagt tcgcaagaag tgatgaaagg aagcgtcata caaagatcca tcttcgtcag
      301 aaggaactag tgaaatctga attggaagag aagaaatctg aacttagaca taaattgaaa
      361 tatgtgccac atgaatatat tgaattgatt gaaatcgcaa gaaattcaac tcaggataga
      421 atccttgaaa tgaaggtgat ggagttcttt atgaaggttt atggttatcg tggtaaacat
      481 ttgggtggat caaggaaacc agacggagca atttatactg tcggatctcc tattgattac
      541 ggtgtgatcg ttgatactaa ggcatattca ggaggttata atcttccaat tggtcaagca
      601 gatgaaatgc aaagatatgt cgaagagaat caaacaagaa acaagcatat caaccctaat
      661 gaatggtgga aagtctatcc atcttcagta acagaattta agttcttgtt tgtgagtggt
      721 catttcaaag gaaactacaa agctcagctt acaagattga atcatatcac taattgtaat
      781 ggagctgttc ttagtgtaga agagcttttg attggtggag aaatgattaa agctggtaca
      841 ttgacacttg aggaagtgag aaggaaattt aataacggtg agataaactt ttaataggac
      901 gtccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
      961 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     1021 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     1081 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     1141 atctatgtta ctagatcggg aattgatccc ccctcgacag cttccggaaa gggcgaattc
     1201 gcaactttgt atacaaaagt tgaacgagaa acgtaaaatg atataaatat caatatatta
     1261 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     1321 ctatgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1381 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1441 tcatcatgcc tcctctggac cagccaggac agaaatgcct cgacttcgct gctacccaag
     1501 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc
     1561 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
     1621 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
     1681 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1741 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     1801 aagttaaaca tcatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     1861 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     1921 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
     1981 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     2041 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     2101 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     2161 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     2221 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     2281 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     2341 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     2401 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
     2461 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     2521 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
     2581 atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg
     2641 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2701 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2761 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2821 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2881 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2941 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3001 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3061 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3121 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3181 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3241 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3301 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3361 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3421 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3481 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3541 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgcgag ccaggaagag
     3601 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3661 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     3721 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     3781 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     3841 ctcgcgttaa cgcttgcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     3901 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
     3961 caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccgaa
     4021 ttcgcccttc acc
//
