LOCUS       pCMV-EB3-mCherry--CMV-LifeAct-GFP        7066 bp    DNA     circular
            
DEFINITION  
ACCESSION   DNA_pCAGIG
VERSION     DNA_pCAGIG.0
KEYWORDS    .

COMMENT
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            <element id="162404060" name="Linker"
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    <setupInfo allowBlunt="true" allowFivePrime="true"
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DISPLAY_SETUP_END|
COMMENT     Backbone pCAGIG Addgene Plasmid #11159
            
            CMV-LifeAct-EGFP was amplified from pCMV-LifeAct-EGFP (Clontech
            backbone, Riedl et al. 2008) with:
            
            CMV-EB3-SpeI FP: 5’-gttactagtagtaatcaattacggggtc-3’
            
            IRES-LA-PA RP 5’-AAAAGATCTCAGTGGTATTTGTGAGCCAGGCCGCTTTACTTGTACAG-3’

            PCR fragment containing CMV-LifeAct-EGFP was digested with
            SpeI/BglII and inserted in a (SpeI/BglII)-digested pCAGIG
            (CAG-IRES-GFP is therefore removed) with in line with the rabbit
            globin polyadenilation signal of pCAGIG resulting in
            pCMV-LifeAct-GFP-pA.

Followed
            
CMV-EB3-mCherry-PA was amplified from pCMV-EB3-mCherry (A.
            Akmanova) with primers
            
            CMV-EB3-SpeI FP: 5’-gttactagtagtaatcaattacggggtc-3’

CMV-EB3-SpeI
            RP: 5’-CTTTACTAGTGGACTCTTGTTCCAAACTGG-3’

Fragment was SpeI digested
            and cloned in a SpeI site preceding the CMV promoter of the above
            described pCMV-LifeAct-GFP-pA vector.

Resistance: Ampicilin
            VNTEXTCHREPL|Animal/Other Eukaryotic
            VNTREPLTYPE|Plasmid
            VNTSEQSTYLE|84|371||0|null|null|null|null|39,48,239
FEATURES             Location/Qualifiers
     polyA_signal    4087..4523
                     /label=rb_globin_polyA_terminator
                     /type="polyA_signal"
                     /vntifkey="25"
     eukaryotic      4753..4932
                     /label=sv40_promoter
                     /promoter_type="eukaryotic"
                     /type="eukaryotic"
                     /vntifkey="29"
     misc_signal     4788..4865
                     /label=sv40_origin
                     /type="misc_signal"
                     /vntifkey="87"
     oriT            5301..5920
                     /label=pBR322_origin
                     /type="oriT"
                     /vntifkey="104"
     misc_feature    complement(6075..6935)
                     /label=Ampicilin Resistance
                     /type="misc_feature"
                     /vntifkey="21"
     promoter        complement(6977..7005)
                     /label=ampicilin promoter
                     /type="promoter"
                     /vntifkey="30"
     polyA_signal    2251..2476
                     /label=sv40_polyA
                     /type="polyA_signal"
                     /vntifkey="25"
     polyA_signal    4946..5077
                     /label=sv40_polyA_late terminator
                     /type="polyA_signal"
                     /vntifkey="25"
     misc_marker     1522..2232
                     /label=mCherry
                     /type="misc_marker"
                     /vntifkey="22"
     mat_peptide     664..1506
                     /label=EB3
                     /note="+TIP End Binding Protein 3"
                     /type="mat_peptide"
                     /vntifkey="84"
     misc_structure  664..2232
                     /label=EB3-mCherry
                     /type="misc_structure"
                     /vntifkey="88"
     eukaryotic      18..615
                     /label=CMV promoter
                     /promoter_type="eukaryotic"
                     /type="eukaryotic"
                     /vntifkey="29"
     misc_marker     3341..4060
                     /label=EGFP
                     /type="misc_marker"
                     /vntifkey="22"
     misc_marker     3269..3319
                     /label=LifeAct
                     /type="misc_marker"
                     /vntifkey="22"
     misc_feature    3320..3340
                     /label=Linker
                     /type="misc_feature"
                     /vntifkey="21"
     misc_feature    3269..4060
                     /label=LifeactGFP
                     /type="misc_feature"
                     /vntifkey="21"
     eukaryotic      2640..3237
                     /label=CMV
                     /promoter_type="eukaryotic"
                     /type="eukaryotic"
                     /vntifkey="29"
ORIGIN
        1 gtcgacattg attattgact agtagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca gtcgactgcc
      661 accatggccg tcaatgtgta ctccacatct gtgaccagtg aaaatctgag tcgccatgat
      721 atgcttgcat gggtcaacga ctccctgcac ctcaactata ccaagataga acagctttgt
      781 tcaggggcag cctactgcca gttcatggac atgctcttcc ccggctgtgt gcacttgagg
      841 aaagtgaagt tccaggccaa actagagcat gaatacatcc acaacttcaa ggtgctgcaa
      901 gcagctttca agaagatggg tgttgacaaa atcattcctg tagagaaatt agtgaaagga
      961 aaattccaag ataattttga gtttattcag tggtttaaga aattctttga cgcaaactat
     1021 gatggaaagg attacaaccc tctgctggcg cggcagggcc aggacgtagc gccacctcct
     1081 aacccaggtg atcagatctt caacaaatcc aagaaactca ttggcacagc agttccacag
     1141 aggacgtccc ccacaggccc aaaaaacatg cagacctctg gccggctgag caatgtggcc
     1201 cccccctgca ttctccggaa gaatcctcca tcagcccgaa atggcggcca tgagactgat
     1261 gcccaaattc ttgaactcaa ccaacagctg gtggacttga agctgacagt ggatgggctg
     1321 gagaaggaac gtgacttcta cttcagcaaa cttcgtgaca tcgagctcat ctgccaggag
     1381 catgaaagtg aaaacagccc tgttatctca ggcatcattg gcatcctcta tgccacagag
     1441 gaaggattcg caccccctga ggacgatgag attgaagagc atcaacaaga agaccaggac
     1501 gagtacctgg atcccgccac catggtgagc aagggcgagg aggataacat ggccatcatc
     1561 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     1621 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     1681 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     1741 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     1801 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1861 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1921 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1981 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     2041 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     2101 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     2161 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     2221 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     2281 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2341 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2401 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2461 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2521 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2581 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2641 ctagtagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     2701 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     2761 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     2821 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     2881 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     2941 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     3001 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     3061 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     3121 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     3181 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
     3241 tcagatctcg agctcaagct tcgaattcat gggtgtcgca gatttgatca agaaattcga
     3301 aagcatctca aaggaagaag gggatccacc ggtcgccacc atggtgagca agggcgagga
     3361 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3421 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3481 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3541 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3601 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3661 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3721 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3781 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3841 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3901 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     3961 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4021 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggcctgg ctcacaaata
     4081 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4141 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4201 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     4261 tttggtttag agtttggcaa catatgccat atgctggctg ccatgaacaa aggtggctat
     4321 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     4381 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     4441 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     4501 cagtcatagc tgtccctctt ctcttatgaa gatccctcga cctgcagccc aagcttggcg
     4561 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4621 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4681 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     4741 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4801 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4861 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4921 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     4981 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5041 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     5101 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     5521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6121 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6181 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6241 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6301 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6361 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6421 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6481 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6541 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6601 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6661 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6721 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6781 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6841 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6901 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
