LOCUS       pLenti6.3-Paxillin-EGFP       10428 bp    DNA     circular UNA 07-MAR-2012
DEFINITION  attB1-attB2-attB5.
ACCESSION   urn.local...iyz-eivu6kb
VERSION     urn.local...iyz-eivu6kb
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     ##Gateway_Cloning_Description-START##
            Design Description :: Mix Entry Clones pE(2F)EGFPpA (L5-L2), pE(2F)
                                  Chicken Paxillin L1R5 and Destination Vector
                                  pLenti6.3/V5-DEST in LR buffer and TE buffers
                                  Thaw, vortex and add LR Clonase enzyme (LR
                                  Clonase Plus enzyme for multi-site Gateway).
                                  Incubate >= 1 hr. at 25�. Add Proteinase K
                                  for 10 min at 37�. Transform into DH5a E.
                                  coli cells (One Shot TOP10 for multi-site
                                  Gateway) or any non-F' episome bearing
                                  strain. Select Expression Clones on agar
                                  plates with antibiotic ampicillin. Express in
                                  appropriate host cells.
            ##Gateway_Cloning_Description-END##
FEATURES             Location/Qualifiers
     misc_feature    67..108
                     /label="V5 epitope"
     misc_feature    127..724
                     /label="WPRE"
     misc_feature    735..1043
                     /label="SV40 promoter"
                     /note="Geneious type: promoter eukaryotic"
     misc_feature    1098..1164
                     /label="EM7 promoter"
                     /note="Geneious type: promoter prokaryotic"
     CDS             1165..1563
                     /label="Blasticidin resistance gene"
     LTR             1649..1883
                     /label="deltaU3"
     LTR             1703..1883
                     /label="3'_LTR"
     polyA_signal    1955..2086
                     /label="SV40 pA"
     misc_feature    2945..3043
                     /label="bla promoter"
                     /note="Geneious type: promoter eukaryotic"
     CDS             3044..3904
                     /label="Amp(R)"
     rep_origin      4049..4722
                     /label="pUC origin"
     misc_feature    5130..5358
                     /label="RSV promoter"
                     /note="Geneious type: promoter eukaryotic"
     LTR             5359..5539
                     /label="HIV-1 5' LTR"
     splicing_signal 5649
                     /label="5' splice donor"
     misc_feature    5650..5694
                     /label="HIC-1 psi packaging signal"
     misc_feature    6204..6437
                     /label="HIV-1 Rev response element (RRE)"
     splicing_signal 6785
                     /label="3' splice acceptor"
     splicing_signal 6813
                     /label="3' splice acceptor"
     misc_feature    6930..7052
                     /label="cPPT"
     misc_feature    7064..7648
                     /label="CMV promoter"
                     /note="Geneious type: promoter eukaryotic"
     attB1           7698..7718
                     /label="attB1"
     CDS             7727..9403
                     /label="Chicken Paxillin"
     modified_base   7817..7819
                     /label="Phosphotyrosine"
     misc_feature    7862..7885
                     /label="Pro-Rich"
     modified_base   8078..8080
                     /label="Phosphotyrosine; by FAK1"
     misc_feature    8702..8854
                     /label="LIM1"
     misc_feature    8879..9031
                     /label="LIM2"
     misc_feature    9056..9208
                     /label="LIM3"
     misc_feature    9233..9385
                     /label="LIM4"
     attB5           9405..9425
                     /label="attB5"
     CDS             9428..10144
                     /label="EGFP"
     misc_structure  9619..9632
                     /label="EGFP-like chromophore"
     misc_feature    10145..10204
                     /label="pCS2 MCS fragment"
     polyA_signal    10221..10416
                     /label="SV40 late polyA (from pCS2+)"
     attB2           complement(10421..13)
                     /label="attB2"
ORIGIN      
        1 ttgtacaaag tggttgatat ccagcacagt ggcggccgct cgagtctaga gggcccgcgg
       61 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggttag
      121 taatgatcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
      181 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
      241 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
      301 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
      361 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
      421 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
      481 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
      541 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
      601 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
      661 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
      721 ctggcgatgg taccggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      781 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
      841 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
      901 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
      961 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     1021 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     1081 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     1141 tacgacaagg tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc
     1201 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     1261 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     1321 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     1381 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     1441 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     1501 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     1561 taagcacaat tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     1621 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     1681 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     1741 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1801 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1861 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     1921 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     1981 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2041 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     2101 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2161 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2221 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     2281 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2341 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     2401 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     2461 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2521 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2581 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2641 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2701 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2761 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2821 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2881 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     2941 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3001 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3061 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3121 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3181 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3241 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3301 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3361 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3421 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3481 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3541 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3601 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3661 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3721 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3781 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3841 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3901 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3961 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4021 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4081 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4141 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4201 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4261 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4321 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4381 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4441 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4501 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4561 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4621 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4681 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4741 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4801 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4861 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     4921 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     4981 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5041 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     5101 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     5161 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     5221 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     5281 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     5341 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5521 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     5581 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     5641 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     5701 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     5761 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     5821 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     5881 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     5941 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     6001 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     6061 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     6121 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     6181 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     6241 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     6301 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     6361 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     6421 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     6481 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     6541 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     6601 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     6661 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     6721 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     6781 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     6841 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     6901 ttagtgaacg gatctcgacg gtatcggtta acttttaaaa gaaaaggggg gattgggggg
     6961 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     7021 caaaaacaaa ttacaaaaat tcaaaatttt atcgataagc ttgggagttc cgcgttacat
     7081 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     7141 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     7201 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     7261 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     7321 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     7381 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     7441 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     7501 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     7561 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
     7621 gctgttttga cctccataga agacaccgac tctagaggat ccactagtcc agtgtggtgg
     7681 aattctgcag atatcaacaa gtttgtacaa aaaagcaggc ttaaccatgg acgacctcga
     7741 tgccttactg gcagacctgg aatccaccac ctcgcatatc tctaagcgac cggtgtttct
     7801 aacagaggaa acgccttact cctacccaac tggaaaccat acgtaccagg agattgctgt
     7861 gccaccgccg gtgcccccgc cgccttccag tgaggcccta aatggcactg tgattgaccc
     7921 cttagaccag tggcaaccca gcgtgtccag atacggccac cagcaacctc agtcccagtc
     7981 tccgatatac agctctagtg ccaaaagctc cagtgcctct gttcctagag acgggctcag
     8041 ctccccttct ccccgtgcca gtgaggagga acacgtgtac agcttcccaa acaagcagaa
     8101 gtctgcagaa ccatctccca caatgaccag cacctccctg ggcagcaacc tctcggaact
     8161 ggacaggctt ctcctggaac tgaatgctgt tcaacataat ccccccagtg gcttctcagc
     8221 agatgaagtc agcagaagcc catcactgcc caatgtgacc ggacctcact atgtcatccc
     8281 agagagcagc agctctgcag gagggaaggc tgcaccccct acaaaagaaa aaccaaagcg
     8341 gaatggtggg cgtgggattg aagatgtgcg tcccagtgtg gagagcctgc tggatgagct
     8401 ggagagctct gtgccaagcc cagtccctgc aatcactgtg agccaagggg aggtgagcag
     8461 cccccagcgt gtcaacgcca gtcagcagca gacccgtatc tctgcttctt cagctacacg
     8521 agaactggat gagctgatgg cgtccctctc tgactttaag ttcatggcac aggggaaagc
     8581 cggcggcagc agctccccac catccaccac cccaaagcct ggcagtcagc tggacaccat
     8641 gctgggaagt ctgcagtctg acctgaacaa actgggggta gcgacggtcg ccaagggtgt
     8701 gtgcggagcc tgcaagaagc cgattgctgg gcaggtagtt acagccatgg ggaagacctg
     8761 gcaccctgag cacttcgtct gcacccactg ccaggaggag attggatcac ggaacttctt
     8821 tgagcgggat gggcagccct actgtgagaa ggactatcac aacctcttct ctcctcgctg
     8881 ctactactgc aatgggccaa tccttgataa agtggtgacg gctttggaca ggacgtggca
     8941 ccctgaacac tttttctgtg cccagtgtgg agttttcttt ggacctgaag gatttcatga
     9001 gaaggatggc aaagcctatt gccgcaagga ctactttgac atgtttgctc ccaagtgtgg
     9061 aggctgtgcc cgggctatcc tggaaaatta catttctgcc ttaaacaccc tgtggcaccc
     9121 tgagtgcttt gtctgtcggg aatgtttcac accgttcatc aacggcagct tcttcgagca
     9181 cgacggccag ccctactgcg aggtgcatta ccacgagcgc cgcggctcgc tgtgctctgg
     9241 ctgccagaag cccatcacgg gacgctgcat cactgccatg ggcaagaaat ttcaccccga
     9301 acactttgtc tgtgccttct gcctcaagca gctcaacaaa ggaaccttca aagaacagaa
     9361 cgacaagccc tactgtcaga actgctttct caaactcttc tgtacaactt tgtatacaaa
     9421 agttgtgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     9481 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     9541 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     9601 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     9661 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     9721 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     9781 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     9841 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     9901 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     9961 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
    10021 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
    10081 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
    10141 caagggcggt ggaagatctg ggaattcaag gcctctcgag cctctagatt ctgcagccct
    10201 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
    10261 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
    10321 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
    10381 tgtttcaggt tcagggggag gtgtgggagg tttttttacc cagctttc
//
