LOCUS       sequence-78692-e        5421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with attL5 and attL2 sites flanking us:NPTII 
            repair template sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5421)
  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 5421)
  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: pNJB98
FEATURES             Location/Qualifiers
     source          1..5421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1475..2266
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1526..1545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2136..2155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      2293..2545
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(2570..2588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(2635..2734)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2752..2770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2775..2791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2775..2791)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3222..4013
                     /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      4106..4694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4595..4614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4848..4865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5024..5051)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5143..5229)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5278..5300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5292..5309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5293..5309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5324..5419
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gagctcgcgg ccgcagcagt cttacttcca tgatttcttt aactatgccg gaatccatcg
       61 cagcgtaatg ctctacacca cgccgaacac ctgggtggac gatatcaccg tggtgacgca
      121 tgtcgcgcaa gactgtaacc acgcgtctgt tgactggcag gtggtggcca atggtgatgt
      181 cagcgttgaa ctgcgtgatg cggatcaaca ggtggttgca actggacaag gcactagcgg
      241 gactttgcaa gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc tctatgaact
      301 gtgcgtcaca gccaaaagcc agacagagtg tgatatctac ccgcttcgcg tcggcatccg
      361 gtcagtggca gtgaagggcg aacagttcct gattaaccac aaaccgttct actttactgg
      421 ctttggtcgt catgaagatg cggacttgcg tggcaaagga ttcgataacg tgctgatggt
      481 gcacgaccac gcattaatgg actggattgg ggccaactcc taccgtacct cgcattaccc
      541 ttacgctgaa gagatgctcg actgggcaga tgaacatggc atcgtggtga ttgatgaaac
      601 tgctgctgtc ggctttaacc tctctttagg cattggtttc gaagcgggca acaagccgaa
      661 agaactgtac agcgaagagg cagtcaacgg ggaaactcag caagcgcact tacaggcgat
      721 taaagagctg atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa
      781 cgaaccggat acccgtccgc aaggtgcacg ggaatatttc gcgccactgg cggaagcaac
      841 gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca
      901 caccgatacc atcagcgatc tctttgatgt gctgtgcctg aaccgttatt acggatggta
      961 tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac ttctggcctg
     1021 gcaggagaaa ctgcatcagc cgattatcat caccgaatac ggcgtggata cgttagccgg
     1081 gctgcactca atgtacaccg acatgtggag tgaagagtat cagtgtgcat ggctggatat
     1141 gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc
     1201 cgattttgcg acctcgcaag gcatattgcg cgttggcggt aacaagaaag ggatcttcac
     1261 tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa
     1321 cttcggtgaa aaaccgcagc agggaggcaa acaacgcagg gaggcaaaca atgatatcac
     1381 aactctcctg acgcgtcatc gtcggctaca gcctcgggaa ttgctaccta gctcgagcaa
     1441 gatccaagga gatataacaa tggcttcctc ctggattgaa caagatggat tgcacgcagg
     1501 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1561 ctgctctgat gccgccgtgt tccggctgtc agcgcagggt agaccggttc tttttgtcaa
     1621 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     1681 ggccacgacg ggcgtacctt gcgctgctgt gctcgacgtt gtcactgaag cgggaaggga
     1741 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     1801 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1861 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1921 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     1981 gttcgccagg ctcaaggcga gaatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2041 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2101 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2161 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2221 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgataac cgcggagagc
     2281 tcgaatttcc ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
     2341 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     2401 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     2461 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     2521 gcggtgtcat ctatgttact agatcgtggg cccgtccagt tccattcgcc ctatagtgag
     2581 tcgtattaaa cctgcattaa tgaatcggcc aacgcgcggg gctgcaggtc gacgacccag
     2641 ctttcttgta caaagttggc attataaaaa ataattgctc atcaatttgt tgcaacgaac
     2701 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     2761 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     2821 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctggacc agccaggaca
     2881 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     2941 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     3001 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     3061 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     3121 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     3181 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg
     3241 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     3301 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     3361 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     3421 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     3481 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     3541 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     3601 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     3661 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     3721 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     3781 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     3841 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     3901 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     3961 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     4021 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     4081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4681 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4741 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4801 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4861 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4921 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4981 ttaatacgcg taccgcgagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     5041 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     5101 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     5161 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     5221 ttttatttga tgcctggcag ttccctactc tcgcgttaac gcttgcatgg atgttttccc
     5281 agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gcccaataat gattttattt
     5341 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
     5401 actttgtata caaaagttgc c
//
