Basic Vector Information
- Vector Name:
- pYB3301
- Antibiotic Resistance:
- Kanamycin
- Length:
- 14707 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Shih PM, Vuu K, Mansoori N, Ayad L, Louie KB, Bowen BP, Northen TR, Loque D.
- Promoter:
- CaMV35S(enhanced)
pYB3301 vector Map
pYB3301 vector Sequence
LOCUS 40924_47513 14707 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pYB3301, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14707)
AUTHORS Shih PM, Vuu K, Mansoori N, Ayad L, Louie KB, Bowen BP, Northen TR,
Loque D.
TITLE A robust gene-stacking method utilizing yeast assembly for plant
synthetic biology
JOURNAL Nat Commun 7, 13215 (2016)
PUBMED 27782150
REFERENCE 2 (bases 1 to 14707)
AUTHORS Shih PM, Vuu K, Mansoori N, Ayad L, Louie K, Bowen BP, Northen TR,
Loque D.
TITLE Direct Submission
JOURNAL Submitted (01-SEP-2016) Feedstocks Division, Joint Bioenergy
Institute, 5885 Hollis St., Emeryville, CA 946082405, USA
REFERENCE 3 (bases 1 to 14707)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14707)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun
7, 13215 (2016)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-SEP-2016) Feedstocks Division, Joint Bioenergy Institute, 5885
Hollis St., Emeryville, CA 946082405, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..14707
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(36..742)
/label=OCS terminator
/note="octopine synthase terminator"
promoter 754..969
/label=URA3 promoter
CDS 970..1770
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
terminator 1857..2107
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 2157..2181
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 3481..4107
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 4544..5608
/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 5677..5871
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 6215..6355
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(6541..7129)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 7385..7792
/label=LEU2 promoter
CDS 7793..8884
/codon_start=1
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
misc_feature complement(9370..9871)
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
CDS complement(9970..10761)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 11186..11210
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
terminator complement(11290..11542)
/label=MAS terminator
/note="mannopine synthase terminator"
CDS complement(11555..12103)
/codon_start=1
/label=BlpR
/note="phosphinothricin acetyltransferase"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(12109..12489)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
promoter 13485..13830
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 14674..14695
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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