LOCUS       Exported               12582 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAL004(Ubi-nos)DII, complete sequence.
ACCESSION   KY313415
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12582)
  AUTHORS   Mir R, Aranda LZ, Baiocchi T, Luo A, Sylvester A, Rasmussen C.
  TITLE     DII-based auxin reporter in maize reveals low auxin signaling during
            telophase and early G1
  JOURNAL   Plant Physiol. (2016) In press
  PUBMED    27881728
REFERENCE   2  (bases 1 to 12582)
  AUTHORS   Rasmussen CG.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-DEC-2016) Botany and Plant Sciences, University of 
            California, Riverside, 900 E. University Ave., Riverside, CA 92521, 
            USA
REFERENCE   3  (bases 1 to 12582)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12582
                     /organism="Cloning vector pAL004(Ubi-nos)DII"
                     /mol_type="other DNA"
                     /db_xref="taxon:1927824"
     misc_feature    complement(95..120)
                     /label=T border(R)
                     /note="Torder(R)"
     misc_feature    115..139
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    297..318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        333..363
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    371..387
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     395..411
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        436..2432
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     regulatory      437..1418
                     /regulatory_class="promoter"
                     /note="Ubi-P"
     misc_feature    2445..2479
                     /label=similar to AtIAA28DII
                     /note="similar to AtIAA28DII"
     CDS             2455..3576
                     /codon_start=1
                     /product="Degron/Venus/N7-NLS"
                     /label=Degron/Venus/N7-NLS
                     /protein_id="APM87885.1"
                     /translation="MKQKSSTKETSFLSNNRVEVAPVVGWPPVRSSRRCGSCDPAFLYK
                     VAAAAAMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGK
                     LPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAE
                     VKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHN
                     IEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGI
                     TLGMDELYIAAAAEFKREEQARKAKVNNEKKTEIVKPESCSNEGDVKDLKRKDSEDGNE
                     GEEEEASSKPKKPKVALSHLQDIDDTEADQEEE"
     misc_feature    2455..2556
                     /label=derived from Degron
                     /note="derived from Degron"
     protein_bind    complement(2570..2594)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2608..3336
                     /label=derived from Venus
                     /note="derived from Venus"
     CDS             2608..3321
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     misc_feature    3337..3576
                     /label=derived from N7-NLS
                     /note="derived from N7-NLS"
     terminator      3599..3851
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      3601..3852
                     /regulatory_class="terminator"
                     /note="NospolyA"
     regulatory      3891..4539
                     /regulatory_class="promoter"
                     /note="35SP"
     promoter        4286..4630
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      4644..4770
                     /regulatory_class="enhancer"
                     /note="TEV"
     CDS             4782..5345
                     /codon_start=1
                     /product="BAR"
                     /label=BAR
                     /note="confers tolerance to herbicide"
                     /protein_id="APM87884.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEICSTI"
     CDS             4782..5330
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     primer_bind     complement(5901..5917)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(6131..6156)
                     /label=T-border(L)
                     /note="T-border(L)"
     misc_feature    6132..6156
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6680..7471
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7717..8305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8491..8631
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      8975..9169
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9235..10308)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(10737..11366)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"