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"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
      661 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
      721 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
      781 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
      841 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
      901 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
      961 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1021 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1081 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1141 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1201 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1261 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1321 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1381 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1441 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaaga tctggatccg
     1501 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     1561 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     1621 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     1681 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     1741 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     1801 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     1861 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     1921 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     1981 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2041 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2101 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2161 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     2221 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     2281 ctttcttgta caaagtggtt ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     2341 cgtagtaatg agctagcgct aaccggtggc gcgttaagtc gacaatcaac ctctggatta
     2401 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2461 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2521 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2581 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2641 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2701 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2761 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     2821 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2881 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2941 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     3001 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     3061 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     3121 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     3181 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     3241 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     3301 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     3361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3481 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3601 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     3661 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     3721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3841 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3901 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4201 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4681 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4981 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6361 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6421 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     6481 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     6541 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     6601 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     6661 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     6721 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     6781 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6841 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6901 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     6961 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     7021 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     7081 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     7141 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     7201 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     7261 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     7321 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     7381 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7441 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     7501 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7561 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7621 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     7681 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7741 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7801 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7861 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7921 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7981 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     8041 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     8101 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     8161 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     8221 tcccaacccc gaggggaccc ttgcgccttt tccaaggcag ccctgggttt gcgcagggac
     8281 gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat
     8341 tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg
     8401 cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga
     8461 cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag
     8521 caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg
     8581 ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg
     8641 ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag
     8701 tcggctccct cgttgaccga atcaccgacc tctctcccca gggggtacca ccggagctta
     8761 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     8821 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     8881 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     8941 acatcggcaa ggtgtgggtc gcggacgacg gcgccgccgt ggcggtctgg accacgccgg
     9001 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     9061 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     9121 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     9181 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     9241 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     9301 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     9361 gagaattcta gatcttgaga caaatggcag tattcatcca caattttaaa agaaaagggg
     9421 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     9481 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     9541 gcagagatcc actttggcgc cggctcgagg ggg
//
