pL1E-lc vector (V005100)

Basic Vector Information

Vector Name:
pL1E-lc
Antibiotic Resistance:
Ampicillin
Length:
4472 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, Mueller-Roeber B.
Promoter:
SNR52

pL1E-lc vector Map

pL1E-lc4472 bp600120018002400300036004200oriAmpRCEN/ARSSNR52 promotergRNA scaffoldSUP4 terminatorCYC1 terminatorI-SceIhomology region E0*TRP1homology region F0*I-SceI

pL1E-lc vector Sequence

LOCUS       40924_27464        4472 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1E-lc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4472)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     AssemblX: a user-friendly toolkit for rapid and reliable multi-gene 
            assemblies
  JOURNAL   Nucleic Acids Res. (2017) In press
  PUBMED    28130422
REFERENCE   2  (bases 1 to 4472)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2016) Department of Molecular Biology - Cell2Fab 
            research unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, 
            Potsdam, Brandenburg 14476, Germany
REFERENCE   3  (bases 1 to 4472)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4472)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-2016) Department of Molecular Biology - Cell2Fab research 
            unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, Potsdam, 
            Brandenburg 14476, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(64..652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(826..1683)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    complement(1781..2284)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        2545..2813
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        2822..2896
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2901..2920
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      2972..3161
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    3380..3397
                     /label=I-SceI
                     /note="I-SceI"
     misc_feature    3398..3447
                     /label=homology region E0*
                     /note="homology region E0*"
     CDS             3456..4127
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     misc_feature    4227..4276
                     /label=homology region F0*
                     /note="homology region F0*"
     misc_feature    4277..4294
                     /label=I-SceI
                     /note="I-SceI"

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