LOCUS       Exported               12347 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUA-At, complete sequence.
ACCESSION   JX992842
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12347)
  AUTHORS   Zhou J, Yu F, Chen B, Wang X, Yang Y, Cheng Y, Yan C, Chen J.
  TITLE     Universal vectors for constructing artificial microRNAs in plants
  JOURNAL   Biotechnol. Lett. 35 (7), 1127-1133 (2013)
  PUBMED    23568377
REFERENCE   2  (bases 1 to 12347)
  AUTHORS   Zhou J, Wang XM, Yan CQ, Chen JP.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-OCT-2012) Zhejiang Provincial Key Laboratory of Plant 
            Virology, Zhejiang Academy of Agricultural Sciences, 198 Shiqiao 
            Road, Hangzhou, Zhejiang 310021, China
REFERENCE   3  (bases 1 to 12347)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12347
                     /organism="Cloning vector pUA-At"
                     /mol_type="other DNA"
                     /db_xref="taxon:1283733"
     regulatory      7..854
                     /regulatory_class="promoter"
                     /note="35S promoter"
     promoter        501..846
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    868..968
                     /label=amiRNA backbone
                     /note="amiRNA backbone"
     promoter        1145..1247
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1248..1928
                     /codon_start=1
                     /gene="Cm(R)"
                     /product="Cm(R)"
                     /label=Cm(R)
                     /protein_id="AGC74796.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     gene            1248..1928
                     /gene="Cm(R)"
                     /label=Cm(R)
     CDS             2248..2553
                     /codon_start=1
                     /gene="ccdB"
                     /product="ccdB"
                     /label=ccdB
                     /protein_id="AGC74795.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2248..2553
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    2620..2774
                     /label=amiRNA backbone
                     /note="amiRNA backbone"
     terminator      2801..3442
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     regulatory      2801..3440
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     primer_bind     complement(3450..3466)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3669..3693
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    3688..3713
                     /label=right T-border
                     /note="right T-border"
     CDS             4993..5622
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6056..7123
                     /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      7189..7383
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7727..7867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8053..8641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8728..9522)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AGC74797.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9947..9972
                     /label=left T-border
                     /note="left T-border"
     misc_feature    9947..9971
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(10022..10237)
                     /label=CaMV35S polyA signal
                     /note="CaMV35S polyA signal"
     polyA_signal    10049..10223
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(10263..11288)
                     /codon_start=1
                     /product="hygromycin resistance protein"
                     /label=hygromycin resistance protein
                     /protein_id="AGC74798.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     regulatory      complement(11324..12093)
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        complement(11356..12033)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    12224..12245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12260..12290
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12298..12314
                     /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     12322..12338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"