LOCUS       40924_34146        8026 bp DNA     circular SYN 18-DEC-2018
DEFINITION  UNVERIFIED: Expression vector PpcleAmiR4-7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8026)
  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 8026)
  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 8026)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8026)
  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..8026
                     /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             3288..4310
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4523..4699
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(4765..4785)
                     /label=attB2
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     promoter        4808..6790
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     terminator      7257..7963
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(7996..8014)
                     /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 gatccctatg aaaaagcctg
     3301 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3361 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3421 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3481 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg
     3541 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     3601 aaaccgaact gcccgctgtt cttcagccgg tcgcggaggc tatggatgcg atcgctgcgg
     3661 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     3721 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     3781 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     3841 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     3901 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     3961 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     4021 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccacgc ctccgggcgt
     4081 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     4141 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     4201 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     4261 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagagtagat
     4321 gccgaccgaa caagagctga tttcgagaac gcctcagcca gcaactcgcg cgagcctagc
     4381 aaggcaaatg cgagagaacg gccttacgct tggtggcaca gttctcgtcc acagttcgct
     4441 aagctcgctc ggctggtcgc gggagaatta attcggtacg ctgaaatcac cagtctctct
     4501 ctacaaatct atctctctct attttctcca taaataatgt gtgagtagtt tcccgataag
     4561 ggaaattagg gttcttatag ggtttcgctc atgtgttgag catataagaa acccttagta
     4621 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     4681 cagtactaaa atccagatcg atgacctgca gacgcgtctc gaggaattcg gtaccccatc
     4741 agatcaaggg tgggcgcgcc gaaccagctt tcttgtacaa agtggtgatc cccctcgaat
     4801 cgaattccgt gcagcgtgac ccggtcgtgc ccctctctag agataatgag cattgcatgt
     4861 ctaagttata aaaaattacc acatattttt tttgtcacac ttgtttgaag tgcagtttat
     4921 ctatctttat acatatattt aaactttact ctacgaataa tataatctat agtactacaa
     4981 taatatcagt gttttagaga atcatataaa tgaacagtta gacatggtct aaaggacaat
     5041 tgagtatttt gacaacagga ctctacagtt ttatcttttt agtgtgcatg tgttctcctt
     5101 tttttttgca aatagcttca cctatataat acttcatcca ttttattagt acatccattt
     5161 agggtttagg gttaatggtt tttatagact aattttttta gtacatctat tttattctat
     5221 tttagcctct aaattaagaa aactaaaact ctattttagt ttttttattt aataatttag
     5281 atataaaata gaataaaata aagtgactaa aaattaaaca aatacccttt aagaaattaa
     5341 aaaaactaag gaaacatttt tcttgtttcg agtagataat gccagcctgt taaacgccgt
     5401 cgacgagtct aacggacacc aaccagcgaa ccagcagcgt cgcgtcgggc caagcgaagc
     5461 agacggcacg gcatctctgt cgctgcctct ggacccctct cgagagttcc gctccaccgt
     5521 tggacttgct ccgctgtcgg catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg
     5581 cacggcaggc ggcctcctcc tcctctcacg gcacggcagc tacgggggat tcctttccca
     5641 ccgctccttc gctttccctt cctcgcccgc cgtaataaat agacaccccc tccacaccct
     5701 ctttccccaa cctcgtgttg ttcggagcgc acacacacac aaccagatct cccccaaatc
     5761 cacccgtcgg cacctccgct tcaaggtacg ccgctcgtcc tccccccccc cccctctcta
     5821 ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac ttctgttcat
     5881 gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta cacggatgcg
     5941 acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt ggggaatcct
     6001 gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt gtttcgttgc
     6061 atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg
     6121 tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt
     6181 tctagatcgg agtagaattc tgtttcaaac tacctggtgg atttattaat tttggatctg
     6241 tatgtgtgtg ccatacatat tcatagttac gaattgaaga tgatggatgg aaatatcgat
     6301 ctaggatagg tatacatgtt gatgcgggtt ttactgatgc atatacagag atgctttttg
     6361 ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc tagatcggag
     6421 tagaatactg tttcaaacta cctggtgtat ttattaattt tggaactgta tgtgtgtgtc
     6481 atacatcttc atagttacga gtttaagatg gatggaaata tcgatctagg ataggtatac
     6541 atgttgatgt gggttttact gatgcatata catgatggca tatgcagcat ctattcatat
     6601 gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa ttattttgat
     6661 cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt tagccctgcc
     6721 ttcatacgct atttatttgc ttggtactgt ttcttttgtc gatgctcacc ctgttgtttg
     6781 gtgttacttc gcccgggttc gaaatcgata agcttgatat cgaattcctg cagccccaaa
     6841 cacacgctcg gacgcatatt acacatgttc atacacttaa tactcgctgt tttgaattga
     6901 tgttttagga atatatatgt agacccggtc ctaatcggct tcattcacag gtcgtgatat
     6961 gattcaatta gcttccgact cattcatcca aataccgagt cgccaaaatt caaactagac
     7021 tcgttaaatg aatgaatgat gcggtagaca aattggatca ttgattctct ttgattgaag
     7081 cggattagga cctggtctct cttttgtatt ccaattttct tgattaatct ttcctgcaca
     7141 aaaacatgct tgatccacta agtgacatat atgctgcctt cgtatatata gttctggtaa
     7201 aattaacatt ttgggtttat ctttatttaa ggcatcgcca tggatcctct agagtcctgc
     7261 tttaatgaga tatgcgagac gcctatgatc gcatgatatt tgctttcaat tctgttgtgc
     7321 acgttgtaaa aaacctgagc atgtgtagct cagatcctta ccgccggttt cggttcattc
     7381 taatgaatat atcacccgtt actatcgtat ttttatgaat aatattctcc gttcaattta
     7441 ctgattgtac cctactactt atatgtacaa tattaaaatg aaaacaatat attgtgctga
     7501 ataggtttat agcgacatct atgatagagc gccacaataa caaacaattg cgttttatta
     7561 ttacaaatcc aattttaaaa aaagcggcag aaccggtcaa acctaaaaga ctgattacat
     7621 aaatcttatt caaatttcaa aaggccccag gggctagtat ctacgacaca ccgagcggcg
     7681 aactaataac gttcactgaa gggaactccg gttccccgcc ggcgcgcatg ggtgagattc
     7741 cttgaagttg agtattggcc gtccgctcta ccgaaagtta cgggcaccat tcaacccggt
     7801 ccagcacggc ggccgggtaa ccgacttgct gccccgagaa ttatgcagca tttttttggt
     7861 gtatgtgggc cccaaatgaa gtgcaggtca aaccttgaca gtgacgacaa atcgttgggc
     7921 gggtccaggg cgaattttgc gacaacatgt cgaggctcag caggaccggc atgcaagcta
     7981 gcttactagt gatattctat agtgtcacct aaatctgcgg ccgctg
//
