LOCUS       pLX301.        9531 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Lentiviral Gateway(R) destination vector with a puromycin resistance
            marker, for strong expression and improved plasmid yield and viral 
            titer.
ACCESSION   .
VERSION     .
KEYWORDS    pLX301.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9531)
  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 9531)
  AUTHORS   Root Lab
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9531)
  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..9531
                     /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"
     misc_feature    2301..2309
                     /label=stop codons
                     /note="stop codons"
     misc_feature    2342..2930
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             3002..3235
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3313..3447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3474..3609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3630..3648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3658..3674)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3816..4271
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4297..4401
                     /label=AmpR promoter
     CDS             4402..5259
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5433..6021
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6309..6330
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6345..6375
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6383..6399
                     /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     6407..6423
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6444..6462
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6490..6716
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6717..6897
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6944..7069
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7562..7795
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7980..8024
                     /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        8199..8699
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             8721..9317
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    9362..9479
                     /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 tagtaatgag ctagcgctaa ccggtggcgc gttaagtcga
     2341 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2401 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2461 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2521 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2581 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2641 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2701 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2761 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2821 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2881 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     2941 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     3001 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     3061 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     3121 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     3181 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     3241 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     3301 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3361 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3421 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     3481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3541 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3601 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     3661 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3721 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3781 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3841 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3901 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3961 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4021 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4081 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4141 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4201 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4261 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4321 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4381 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4441 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4501 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4561 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4621 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4681 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4741 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4801 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4861 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4921 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4981 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5041 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5101 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5161 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5221 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5281 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5341 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5401 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5461 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5521 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5581 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5641 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5701 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5761 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5821 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5881 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5941 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6001 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6061 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6121 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6181 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6241 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6301 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6361 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6421 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     6481 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     6541 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     6601 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     6661 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     6721 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6781 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6841 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     6901 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     6961 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     7021 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     7081 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     7141 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     7201 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     7261 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     7321 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     7381 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     7441 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     7501 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     7561 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     7621 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     7681 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     7741 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     7801 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     7861 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     7921 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     7981 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     8041 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     8101 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     8161 tatcgtttca gacccacctc ccaaccccga ggggaccctt gcgccttttc caaggcagcc
     8221 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
     8281 accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta
     8341 cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc
     8401 ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag
     8461 acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg
     8521 gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt
     8581 gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc
     8641 cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg
     8701 gggtaccacc ggagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     8761 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     8821 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     8881 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgccgtgg
     8941 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     9001 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     9061 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     9121 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     9181 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     9241 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     9301 cccgcaagcc cggtgcctga gaattctaga tcttgagaca aatggcagta ttcatccaca
     9361 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     9421 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     9481 gggtttatta cagggacagc agagatccac tttggcgccg gctcgagggg g
//
