pLX302 vector (V010410)

LOCUS       pLX302.        9573 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Lentiviral Gateway(R) destination vector with a puromycin resistance
            marker and a V5 epitope tag, for strong expression and improved 
            plasmid yield and viral titer.
ACCESSION   .
VERSION     .
KEYWORDS    pLX302
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9573)
  AUTHORS   Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja 
            R, Thomas SR, Alkan O, Bhimdi T, Green TM, Johannessen CM, Silver 
            SJ, Nguyen C, Murray RR, Hieronymus H, Balcha D, Fan C, Lin C, 
            Ghamsari L, Vidal M, Hahn WC, Hill DE, Root DE.
  TITLE     A public genome-scale lentiviral expression library of human ORFs.
  JOURNAL   Nat. Methods 2011;8:659-61.
  PUBMED    21706014
REFERENCE   2  (bases 1 to 9573)
  AUTHORS   Root Lab
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9573)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2011"; volume: "8"; pages: "659-61"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    595..719
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        744..774
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             828..1484
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1829..2131
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2175..2299)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2301..2342
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     misc_feature    2343..2351
                     /label=stop codons
                     /note="stop codons"
     misc_feature    2384..2972
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             3044..3277
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3355..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3516..3651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3672..3690)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3700..3716)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3858..4313
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4339..4443
                     /label=AmpR promoter
     CDS             4444..5301
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5475..6063
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6351..6372
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6387..6417
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6425..6441
                     /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     6449..6465
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6486..6504
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6532..6758
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6759..6939
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6986..7111
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7604..7837
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8022..8066
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     promoter        8241..8741
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             8763..9359
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    9404..9521
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"