LOCUS       sequence-146016-        8269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit for kanamycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8269)
  AUTHORS   Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo 
            P, Blanca J, Granell A, Orzaez D
  TITLE     A modular toolbox for gRNA-Cas9 genome engineering in plants based 
            on the GoldenBraid standard.
  JOURNAL   Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. 
            eCollection 2016.
  PUBMED    26839579
REFERENCE   2  (bases 1 to 8269)
  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: "Plant 
            Methods."; date: "2016-02-1"; volume: "12:10. doi"; pages: " 
            10.1186/s13007-016-0101-2. eCollection 2016"
COMMENT     Sequence Label: pEGB3omega1R Tnos:NptII:Pnos-SF (GB1181)
FEATURES             Location/Qualifiers
     source          1..8269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(20..272)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(312..1106)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(423..442)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1033..1052
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1140..1319)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    1581..1605
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2906..3535
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3969..5036
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      5102..5296
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5532..5554)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5640..5780
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5695..5714
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5795..5812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5966..6554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6046..6065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6642..7433)
                     /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"
     misc_feature    8219..8243
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 atcagcttgc atgccggtcg atctagtaac atagatgaca ccgcgcgcga taatttatcc
       61 tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta
      121 atcataaaaa cccatctcat aaataacgtc atgcattaca tgttaattat tacatgctta
      181 acgtaattca acagaaatta tatgataatc atggcaagac cggcaacagg attcaatctt
      241 aagaaacttt attgccaaat gtttgaacga tctgcttgac tctagctaga gtccgaaccc
      301 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggctatgcg ctgcgaatcg
      361 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
      421 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
      481 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcgtcg
      541 ccgtgggtca cgacgagatc ctcgccgtcg ggcatccgcg ccttgagcct ggcgaacagt
      601 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
      661 tccatccgag tacgtgctcg ctcgatgcgg tgtttcgctt ggtggtcgaa tgggcaggta
      721 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
      781 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
      841 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
      901 cacgatagcc gcgctgcctc gtcttggagt tcattcaggg caccggacag gtcggtcttg
      961 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     1021 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     1081 gcgtgcaatc catcttgttc aatcatgcct cgatcgagtt gagagtgaat atgagactct
     1141 aattggatac cgaggggaat ttatggaacg tcagtggagc atttttgaca agaaatattt
     1201 gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt
     1261 gcttagctca ttaaactcca gaaacccgcg gctcagtggc tccttcaacg ttgcggttct
     1321 gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat accgtgactc
     1381 ccttaattct catgtatctc ctgacagcga aatgattgat gaagaacaat ggtggatgaa
     1441 gaacaaagaa ggagggagct tttgttcaag atgaacaaag aacaatagtg gatgaagaac
     1501 aaagtgaaaa aaataaaaaa aaatgtatgg ttaaataaag agtaaagtta ccattgagac
     1561 tccgtcatga gaccgaattc tgacaggata tattggcggg taaacctaag agaaaagagc
     1621 gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt
     1681 gtatgtgcat gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc
     1741 cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcatctga aaacgacatg
     1801 tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt
     1861 gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc
     1921 atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag
     1981 gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag
     2041 aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc
     2101 cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta
     2161 ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg
     2221 tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc
     2281 gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc
     2341 cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc
     2401 gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat
     2461 cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg
     2521 cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag
     2581 aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt
     2641 tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg
     2701 cgcacctctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg
     2761 cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat
     2821 gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatc cgatgagtaa
     2881 ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg
     2941 gtcaggcaag acgaccatcg gaacccatct agcccgcgcc ctgcaactcg ccggggccga
     3001 tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga
     3061 agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc
     3121 catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt
     3181 gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat
     3241 atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag
     3301 gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt
     3361 tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt
     3421 gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga
     3481 cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa
     3541 tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca
     3601 cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc
     3661 aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc
     3721 aagaccatta ccgagctgct atctgaatag atcgcgcagc taccagagta aatgagcaaa
     3781 tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca
     3841 accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt
     3901 aagcggctgg gttgtctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc
     3961 ggcgtgacgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
     4021 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
     4081 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
     4141 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
     4201 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
     4261 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
     4321 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
     4381 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
     4441 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
     4501 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
     4561 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
     4621 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
     4681 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
     4741 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
     4801 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
     4861 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
     4921 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
     4981 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
     5041 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggactctttc
     5101 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
     5161 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
     5221 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
     5281 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
     5341 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
     5401 gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc
     5461 cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat
     5521 gacggtgaaa acctctgaca catgcagctc ccggtgacgg tcacagcttg tctgtaagcg
     5581 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5641 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     5701 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     5761 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5821 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5881 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5941 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6601 aaaactcacg ttaagggatt ttggtcatgc attctaggtg attatttgcc gactaccttg
     6661 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     6721 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     6781 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     6841 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc accagcccag
     6901 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     6961 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     7021 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     7081 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     7141 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     7201 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     7261 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     7321 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     7381 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct cataatgttt
     7441 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     7501 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacccca
     7561 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     7621 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag cttacgaacc
     7681 gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     7741 gcaaccttgg gtagcagcga agtcgaggca tttctgtcct ggctggaaca gaacttatta
     7801 tttccttcct cttttctaca gtatttaaag ataccccaag aagctaatta taacaagacg
     7861 aactccaatt cactgttcct tgcattctaa aaccttaaat accagaaaac agctttttca
     7921 aagttgtttt caaagttggc gtataacata gtatcgacgg agccgatttt gaaaccgcgg
     7981 tgatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc
     8041 atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc ggtaacatga
     8101 gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact gatgggctgc
     8161 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg
     8221 gcaggatata ttgtggtgta aacataacga attcggtctc aggagagcg
//
