LOCUS       sequence-156992-        7307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  binary DEST vector for plant expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7307)
  AUTHORS   Jakoby M, Wang HY, Reidt W, Weisshaar B, Bauer P
  TITLE     FRU (BHLH029) is required for induction of iron mobilization genes 
            in Arabidopsis thaliana.
  JOURNAL   FEBS Lett. 2004 Nov 19;577(3):528-34.
  PUBMED    15556641
REFERENCE   2  (bases 1 to 7307)
  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: "FEBS 
            Lett."; date: "2004-11-19"; volume: "577(3)"; pages: "528-34"
COMMENT     Sequence Label: pLEELA
FEATURES             Location/Qualifiers
     source          1..7307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    364..488
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(513..518,519..536,537..543)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             597..1256
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(663..682)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1598..1903
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1782..1801
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1944..2068)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2385..2409
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      2521..3230
                     /label=oriV
                     /note="incP origin of replication"
     promoter        3291..3395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3396..3464,3465..4256)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3614..3633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4427..5015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4916..4935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5327..5351
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(5445..5697)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(5717..6268)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(6310..6489)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        6961..7306
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     7250..7270
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 tcgagaaaga ggatccacct gaggcctccg ttccatgggc tagaagcttc tcctcctctg
       61 ctaacgtaag cctctctgtt ttttttctct gtttcttttg aaatgaatcc aattagtgat
      121 gataatctgt gtttgatgta tcattgattt aacatcttga caatgaatcg tgatcggaag
      181 tgataaagtt atgggtcaac ggtttcaaag agagagaaag acttttagag tcaactctcg
      241 actctttctt aattatgtta ttgctatttg tctcttttct tgaagtctga acaattcttg
      301 ggattgtttt gcaggttcta gcttctccaa ccacaggaat tcatcgcccg ggactgggtt
      361 atcacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata
      421 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
      481 tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata
      541 atgtgtggat tttgagttag gatccggcga gattttcagg agctaaggaa gctaaaatgg
      601 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
      661 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
      721 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
      781 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
      841 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
      901 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
      961 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     1021 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     1081 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     1141 aggtgctgat gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg
     1201 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaacgc
     1261 gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     1321 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt gtgctatgaa
     1381 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     1441 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     1501 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     1561 aacggctctt ttgctgacga gaacagggac tggtgaaatg cagtttaagg tttacaccta
     1621 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     1681 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     1741 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     1801 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     1861 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     1921 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
     1981 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     2041 ctcgttcagc tttcttgtac aaagtggtga taacttccga tcgattctag actagttcta
     2101 gagtccgcaa aaatcaccag tctctctcta caaatctatc tctctctatt tttctccaga
     2161 ataatgtgtg agtagttccc agataaggga attagggttc ttatagggtt tcgctcatgt
     2221 gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa
     2281 aatttctaat tcctaaaacc aaaatccagt gaccgggcgg ccgccaccgc ggtggagggg
     2341 gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag gatatattgg
     2401 cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga
     2461 aaaggtttat ccgttcgtcc atttgtatgt gtacatcacc gacgagcaag gcaagaccga
     2521 gcgcctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
     2581 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgg ccgccggcgt
     2641 tgtggatacc tcgcggaaaa cttggccctc actgacagat gaggggcgga cgttgacact
     2701 tgaggggccg actcacccgg cgcggcgttg acagatgagg ggcaggctcg atttcggccg
     2761 gcgacgtgga gctggccagc ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc
     2821 ccacagatga tgtggacaag cctggggata agtgccctgc ggtattgaca cttgaggggc
     2881 gcgactactg acagatgagg ggcgcgatcc ttgacacttg aggggcagag tgctgacaga
     2941 tgaggggcgc acctattgac atttgagggg ctgtccacag gcagaaaatc cagcatttgc
     3001 aagggtttcc gcccgttttt cggccaccgc taacctgtct tttaacctgc ttttaaacca
     3061 atatttataa accttgtttt taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg
     3121 aaggggggtg cccccccttc tcgaaccctc ccggcccgct aacgcgggcc tcccatcccc
     3181 ccaggggctg cgcccctcgg ccgcgaacgg cctcacccca aaaatggcag cgctggcagt
     3241 ccttgccatt gccgggatcg gggcagtaac gggatgggcg atcagcccga caagctaccc
     3301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3361 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3421 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3481 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3541 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3601 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3661 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3721 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3781 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3841 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3901 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3961 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4021 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4081 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4141 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4201 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4261 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4321 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4381 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4441 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4501 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4561 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4621 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4681 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4741 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4801 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4861 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4921 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4981 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5041 ggttcctggc cttttgctgg ccttttgctc acatggactc tagctagagg atcacaggca
     5101 gcaacgctct gtcatcgtta caatcaacat gctaccctcc gcgagatcat ccgtgtttca
     5161 aacccggcag cttagttgcc gttcttccga atagcatcgg taacatgagc aaagtctgcc
     5221 gccttacaac ggctctcccg ctgacgccgt cccggactga tgggctgcct gtatcgagtg
     5281 gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt
     5341 gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg tttttaatga
     5401 tcgaatacta acgtctctac cagatatcag cttgcatgcc ggtcgatcta gtaacataga
     5461 tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta
     5521 ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca
     5581 ttacatgtta attattacat gcttaacgta attcaacaga aattatatga taatcatcgc
     5641 aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg aacgatctgc
     5701 ttgactctag gggtcatcag atttcggtga cgggcaggac cggacggggc ggcaccggca
     5761 ggctgaagtc cagctgccag aaacccacgt catgccagtt cccgtgcttg aagccggccg
     5821 cccgcagcat gccgcggggg gcatatccga gcgcctcgtg catgcgcacg ctcgggtcgt
     5881 tgggcagccc gatgacagcg accacgctct tgaagccctg tgcctccagg gacttcagca
     5941 ggtgggtgta gagcgtggag cccagtcccg tccgctggtg gcggggggag acgtacacgg
     6001 tcgactcggc cgtccagtcg taggcgttgc gtgccttcca gggacccgcg taggcgatgc
     6061 cggcgacctc gccgtccacc tcggcgacga gccagggata gcgctcccgc agacggacga
     6121 ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg gaagttgacc gtgcttgtct
     6181 cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg gcacggcgga
     6241 tgtcggccgg gcgtcgttct gggctcatgg tagatcccct cgatcgagtt gagagtgaat
     6301 atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc atttttgaca
     6361 agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc
     6421 tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctcagtggc tccttcaacg
     6481 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
     6541 aacgtgactc ccttaattct ccgctcatga tcagattgtc gtttcccgcc ttcggtttgg
     6601 gcgcgcccgg tctcagaaga ccagagggct attgagactt ttcaacaaag ggtaatatcg
     6661 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa
     6721 aaggaagatg gcttctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagaa
     6781 tgcctctacc gacagtggtc ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa
     6841 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgataaca tggtggagca
     6901 cgacactctc gtctactcca agaatatcaa agatacagtc tcagaagacc agagggctat
     6961 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat
     7021 ctgtcacttc atcgaaagga cagtagaaaa ggaagatggc ttctacaaat gccatcattg
     7081 cgataaagga aaggctatcg ttcaagaatg cctctaccga cagtggtccc aaagatggac
     7141 ccccacccac gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     7201 tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
     7261 aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacc
//
