pUA-At vector (V002482)

Basic Vector Information

Vector Name:
pUA-At
Antibiotic Resistance:
Kanamycin
Length:
12347 bp
Type:
Cloning vector
Replication origin:
ori
Host:
Plants
Source/Author:
Zhou J, Yu F, Chen B, Wang X, Yang Y, Cheng Y, Yan C, Chen J.
Promoter:
CaMV35S(long)

pUA-At vector Vector Map

pUA-At12347 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000CaMV 35S promoteramiRNA backbonecat promoterCmRccdBamiRNA backboneE9 terminatorM13 fwdRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanRLB T-DNA repeatCaMV poly(A) signalHygRCaMV 35S promoter (enhanced)CAP binding sitelac promoterlac operatorM13 rev

pUA-At vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_44664       12347 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUA-At, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12347)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Lett."; date: "2013"; volume: "35"; issue: "7"; pages: "1127-1133"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-OCT-2012) Zhejiang Provincial Key Laboratory of Plant Virology, 
            Zhejiang Academy of Agricultural Sciences, 198 Shiqiao Road, 
            Hangzhou, Zhejiang 310021, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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..1925
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             2248..2550
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     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"
     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"
     CDS             4993..5619
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6056..7120
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /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(8731..9522)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9947..9971
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(10049..10223)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(10266..11288)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     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
                     /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
                     /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"

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