LOCUS       40924_34141        7676 bp DNA     circular SYN 18-DEC-2018
DEFINITION  UNVERIFIED: Expression vector PpcleAmiR1-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7676)
  AUTHORS   Whitewoods CD, Cammarata J, Nemec Venza Z, Sang S, Crook AD, Aoyama 
            T, Wang XY, Waller M, Kamisugi Y, Cuming AC, Szovenyi P, Nimchuk ZL,
            Roeder AH, Scanlon MJ, Harrison CJ.
  TITLE     CLAVATA Was a Genetic Novelty for the Morphological Innovation of 3D
            Growth in Land Plants
  JOURNAL   Curr. Biol. 28 (15), 2365-2376 (2018)
  PUBMED    30033333
REFERENCE   2  (bases 1 to 7676)
  AUTHORS   Whitewoods CD, Cammarata J, Nemec Venza Z, Sang S, Crook AD, Aoyama 
            T, Waller M, Kamisugi Y, Cuming AC, Szovenyi P, Nimchuk ZL, Roeder 
            AHK., Scanlon MJ, Harrison CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-APR-2018) Biological Sciences, University of Bristol, 
            24 Tyndall Avenue, Bristol BS81TQ, UK
REFERENCE   3  (bases 1 to 7676)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7676)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr. 
            Biol."; date: "2018"; volume: "28"; issue: "15"; pages: "2365-2376"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-APR-2018) Biological Sciences, University of Bristol, 24 Tyndall
            Avenue, Bristol BS81TQ, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            GenBank staff is unable to verify sequence and/or annotation 
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..7676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..33)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(213..641)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        668..772
                     /label=AmpR promoter
     CDS             773..1630
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1804..2392
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2680..2701
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2716..2746
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2754..2770
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2778..2794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2825..2843
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2926..3271
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3297..4088
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4193..4369
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(4434..4454)
                     /label=attB2
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     promoter        4477..6440
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     terminator      6907..7613
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(7646..7664)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 accaagtcag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
       61 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      121 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggaaattg
      181 taaacgttaa tattttgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
      241 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      301 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      361 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      421 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      481 gatgccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      541 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      601 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
      661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      781 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      841 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
      901 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      961 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1021 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1081 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1141 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1201 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1261 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1321 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1381 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1441 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1501 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1561 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1621 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1681 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1741 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1801 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1861 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1921 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     1981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2161 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2581 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2641 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     2701 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     2761 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgaatttg gccaagtcgg
     2821 cctctaatac gactcactat agggagctcg cggccgcatg catatgtcat cgtaccccta
     2881 ctccaaaaat gtcaaagata cagtctcaga agaccaaagg gctattgaga cttttcaaca
     2941 aagggtaatt tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga
     3001 aaggacagta gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc
     3061 tatcattcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
     3121 catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgacat
     3181 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     3241 ataaggaagt tcatttcatt tggagaggac agcccaagct ggcatgattg aacaagatga
     3301 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3361 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3421 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactccaag
     3481 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3541 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3601 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3661 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3721 agcgaggacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3781 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcggatg cccgacggcg
     3841 aggatctcgt cgtgacccag ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3901 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3961 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4021 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4081 agttcttctg agcgggactc tgtgcgggac tctggggttc ggactctagc tagaggatca
     4141 attcggtacg ctgaaatcac cagtctctct ctacaaatct atctctctct attttctcca
     4201 taaataatgt gtgagtagtt tcccgataag ggaaattagg gttcttatag ggtttcgctc
     4261 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     4321 ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcg atgacctgca
     4381 gacgcgtctc gaggaattcg gtacccatca gatcaagggt gggcgcgccg aaccagcttt
     4441 cttgtacaaa gtggtgatcc ccctcgaatc gaattccgtg cagcgtgacc cggtcgtgcc
     4501 cctctctaga gataatgagc attgcatgtc taagttataa aaaattacca catatttttt
     4561 ttgtcacact tgtttgaagt gcagtttatc tatctttata catatattta aactttactc
     4621 tacgaataat ataatctata gtactacaat aatatcagtg ttttagagaa tcatataaat
     4681 gaacagttag acatggtcta aaggacaatt gagtattttg acaacaggac tctacagttt
     4741 tatcttttta gtgtgcatgt gttctccttt ttttttgcaa atagcttcac ctatataata
     4801 cttcatccat tttattagta catccattta gggtttaggg ttaatggttt ttatagacta
     4861 atttttttag tacatctatt ttattctatt ttagcctcta aattaagaaa actaaaactc
     4921 tattttagtt tttttattta ataatttaga tataaaatag aataaaataa agtgactaaa
     4981 aattaaacaa atacccttta agaaattaaa aaaactaagg aaacattttt cttgtttcga
     5041 gtagataatg ccagcctgtt aaacgccgtc gacgagtcta acggacacca accagcgaac
     5101 cagcagcgtc gcgtcgggcc aagcgaagca gacggcacgg catctctgtc gctgcctctg
     5161 gacccctctc gagagttccg ctccaccgtt ggacttgctc cgctgtcggc atccagaaat
     5221 tgcgtggcgg agcggcagac gtgagccggc acggcaggcg gcctcctcct cctctcacgg
     5281 cacggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc ctcgcccgcc
     5341 gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt tcggagcgca
     5401 cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt caaggtacgc
     5461 cgctcgtcct cccccccccc ccctctctac cttctctaga tcggcgttcc ggtccatggt
     5521 tagggcccgg tagttctact tctgttcatg tttgtgttag atccgtgttt gtgttagatc
     5581 cgtgctgcta gcgttcgtac acggatgcga cctgtacgtc agacacgttc tgattgctaa
     5641 cttgccagtg tttctctttg gggaatcctg ggatggctct agccgttccg cagacgggat
     5701 cgatttcatg attttttttg tttcgttgca tagggtttgg tttgcccttt tcctttattt
     5761 caatatatgc cgtgcacttg tttgtcgggt catcttttca tgcttttttt tgtcttggtt
     5821 gtgatgatgt ggtctggttg ggcggtcgtt ctagatcgga gtagaattct gtttcaaact
     5881 acctggtgga tttattaatt ttggatctgt atgtgtgtgc catacatatt catagttacg
     5941 aattgaagat gatggatgga aatatcgatc taggataggt atacatgttg atgcgggttt
     6001 tactgatgca tatacagaga tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg
     6061 ggcggtcgtt cattcgttct agatcggagt agaatactgt ttcaaactac ctggtgtatt
     6121 tattaatttt ggaactgtat gtgtgtgtca tacatcttca tagttacgag tttaagatgg
     6181 atggaaatat cgatctagga taggtataca tgttgatgtg ggttttactg atgcatatac
     6241 atgatggcat atgcagcatc tattcatatg ctctaacctt gagtacctat ctattataat
     6301 aaacaagtat gttttataat tattttgatc ttgatatact tggatgatgg catatgcagc
     6361 agctatatgt ggattttttt agccctgcct tcatacgcta tttatttgct tggtactgtt
     6421 tcttttgtcg atgctcaccc gggttcgaaa tcgataagct tgatatcgaa ttcctgcagc
     6481 cccaaacaca cgctcggacg catattacac atgttcatac acttaatact cgctgttttg
     6541 aattgatgtt ttaggaatat atatgtagac tacgaccgca tgcttcccat tcacaggtcg
     6601 tgatatgatt caattagctt ccgactcatt catccaaata ccgagtcgcc aaaattcaaa
     6661 ctagactcgt taaatgaatg aatgatgcgg tagacaaatt ggatcattga ttctctttga
     6721 ttgggaacca tgcggtcgga gtctctcttt tgtattccaa ttttcttgat taatctttcc
     6781 tgcacaaaaa catgcttgat ccactaagtg acatatatgc tgccttcgta tatatagttc
     6841 tggtaaaatt aacattttgg gtttatcttt atttaaggca tcgccatggg gggatcctct
     6901 agagtcctgc tttaatgaga tatgcgagac gcctatgatc gcatgatatt tgctttcaat
     6961 tctgttgtgc acgttgtaaa aaacctgagc atgtgtagct cagatcctta ccgccggttt
     7021 cggttcattc taatgaatat atcacccgtt actatcgtat ttttatgaat aatattctcc
     7081 gttcaattta ctgattgtac cctactactt atatgtacaa tattaaaatg aaaacaatat
     7141 attgtgctga ataggtttat agcgacatct atgatagagc gccacaataa caaacaattg
     7201 cgttttatta ttacaaatcc aattttaaaa aaagcggcag aaccggtcaa acctaaaaga
     7261 ctgattacat aaatcttatt caaatttcaa aaggccccag gggctagtat ctacgacaca
     7321 ccgagcggcg aactaataac gttcactgaa gggaactccg gttccccgcc ggcgcgcatg
     7381 ggtgagattc cttgaagttg agtattggcc gtccgctcta ccgaaagtta cgggcaccat
     7441 tcaacccggt ccagcacggc ggccgggtaa ccgacttgct gccccgagaa ttatgcagca
     7501 tttttttggt gtatgtgggc cccaaatgaa gtgcaggtca aaccttgaca gtgacgacaa
     7561 atcgttgggc gggtccaggg cgaattttgc gacaacatgt cgaggctcag caggaccggc
     7621 atgcaagcta gcttactagt gatattctat agtgtcacct aaatctgcgg ccgctg
//
