LOCUS       Exported                6615 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses two mutated tandem repeats of Tapp1 PH domain 
            (phosphoinositide binding-deficient mutant of TAPP1, R211L, can't 
            bind PI(3,4)P2). Fused to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    GFP-2xPH-TAPP1 R211L mutant
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  AUTHORS   Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, 
            McGarry L, Shanks E, Mostov KE, Bryant DM
  TITLE     The phospholipid PI(3,4)P2 is an apical identity determinant.
  JOURNAL   Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8.
  PUBMED    30487552
REFERENCE   2  (bases 1 to 6615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6615
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        96..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     554..574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      642..651
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             648..1364
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             648..1364
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(693..714)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(954..973)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1301..1322
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2337..2346
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2346..2375
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3438..3559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3475..3494)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3529..3548
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3566..4021)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3703..3724)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3915..3934
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4048..4152
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4154..4511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4177..4197)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4362..4497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4424..4443
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4546..5340
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4600..4619)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5210..5229
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5528..5547)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5572..5619
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5948..6536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6437..6456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
       61 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      121 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      181 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      241 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      301 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      361 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      421 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      481 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      541 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      601 gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg
      661 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      721 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      781 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
      841 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
      901 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      961 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1021 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1081 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1141 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1201 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1261 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1321 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc
     1381 gagtgaatga tcagcaggac ctagtggaat gggtaaatgt gttaaacaaa gctataaaaa
     1441 ttacagtacc aaagcagtca gactcacagc ctaattctga taacctaagt cgccatggtg
     1501 aatgtgggaa aaagcaagtg tcttacagaa ctgatattgt tggtggcgta cccatcatta
     1561 ctcccactca gaaagaagaa gtaaatgaat gtggtgaaag tattgacaga aataatctga
     1621 aacggtcaca aagccatctt ccttacttta ctcctaaacc acctcaagat agtgcggtta
     1681 tcaaagctgg atattgtgta aaacaaggag cagtgatgaa aaactggaag agactgtatt
     1741 ttcaattgga tgaaaacaca ataggctact tcaaatctga actggaaaag gaacctcttc
     1801 gcgtaatacc acttaaagag gttcataaag tccaggaatg taagcaaagc gacataatga
     1861 tgagggacaa cctctttgaa attgtaacaa cgtctcgaac tttctatgtg caggctgata
     1921 gccctgaaga gatgcacagt tggattaaag cagtctctgg cgccattgta gcacagcggg
     1981 gtcccggcag atctgcgtct tctgagcatc cccccggtcc ttcagaatcc aaacacgctt
     2041 tccgtcctac caacgcagcc gccgccacct cacattccac agcctctcgc agcaactctt
     2101 tggtctcaac ctttaccatg gagaagcgag gattttacga gtctcttgcc aaggtcaagc
     2161 cagggaactt caaggtccag actgtctctc caagagaacc agcttccaaa gtgactgaac
     2221 aagctctgtt aagacctcaa agtaaaaatg gccctcagga aaaagattgt gacctagtag
     2281 acttggacga tgcgagcctt ccggtcagtg acgtgggtac cgcgggcccg ggatccgcca
     2341 ccatggaaca aaaactcatc tcagaagagg atctgaatga tcagcaggac ctagtggaat
     2401 gggtaaatgt gttaaacaaa gctataaaaa ttacagtacc aaagcagtca gactcacagc
     2461 ctaattctga taacctaagt cgccatggtg aatgtgggaa aaagcaagtg tcttacagaa
     2521 ctgatattgt tggtggcgta cccatcatta ctcccactca gaaagaagaa gtaaatgaat
     2581 gtggtgaaag tattgacaga aataatctga aacggtcaca aagccatctt ccttacttta
     2641 ctcctaaacc acctcaagat agtgcggtta tcaaagctgg atattgtgta aaacaaggag
     2701 cagtgatgaa aaactggaag agactgtatt ttcaattgga tgaaaacaca ataggctact
     2761 tcaaatctga actggaaaag gaacctcttc gcgtaatacc acttaaagag gttcataaag
     2821 tccaggaatg taagcaaagc gacataatga tgagggacaa cctctttgaa attgtaacaa
     2881 cgtctcgaac tttctatgtg caggctgata gccctgaaga gatgcacagt tggattaaag
     2941 cagtctctgg cgccattgta gcacagcggg gtcccggcag atctgcgtct tctgagcatc
     3001 cccccggtcc ttcagaatcc aaacacgctt tccgtcctac caacgcagcc gccgccacct
     3061 cacattccac agcctctcgc agcaactctt tggtctcaac ctttaccatg gagaagcgag
     3121 gattttacga gtctcttgcc aaggtcaagc cagggaactt caaggtccag actgtctctc
     3181 caagagaacc agcttccaaa gtgactgaac aagctctgtt aagacctcaa agtaaaaatg
     3241 gccctcagga aaaagattgt gacctagtag acttggacga tgcgagcctt ccggtcagtg
     3301 acgtgtgagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3361 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3421 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3481 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3541 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3601 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3661 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3721 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3781 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3841 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3901 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3961 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4021 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4081 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4141 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4201 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4261 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4381 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4441 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4501 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4561 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4621 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4681 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4741 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4801 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4861 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4921 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4981 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5041 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5101 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5161 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5221 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5281 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5341 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5401 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5461 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5521 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5581 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5641 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5701 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5761 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5821 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5881 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5941 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6001 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6061 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6121 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6181 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6241 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6301 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6361 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6421 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6481 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6541 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6601 ttcctgcgtt atccc
//
