LOCUS       sequence-501957-        7857 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Expresses two tandem repeats of Tapp1 PH domain that binds to 
            PI(3,4)P2. Fused to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  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 7857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-11-28"; volume: "9(1):5041. doi"; pages: " 
            10.1038/s41467-018-07464-8"
COMMENT     Sequence Label: GFP-2xPH-TAPP1 WT
FEATURES             Location/Qualifiers
     source          1..7857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2302..2311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2311..2340
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3412..3421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3421..3450
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    4645..4766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4682..4701)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4736..4755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4773..5228)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4910..4931)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5122..5141
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5255..5359
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5361..5718
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5361..5552
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5384..5404)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5569..5704
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5753..6547
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5753..6547
                     /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(5807..5826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6417..6436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6735..6754)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6779..6826
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7155..7743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7644..7663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagtg aatgatcagc aggacctagt ggaatgggta
     1381 aatgtgttaa acaaagctat aaaaattaca gtaccaaagc agtcagactc acagcctaat
     1441 tctgataacc taagtcgcca tggtgaatgt gggaaaaagc aagtgtctta cagaactgat
     1501 attgttggtg gcgtacccat cattactccc actcagaaag aagaagtaaa tgaatgtggt
     1561 gaaagtattg acagaaataa tctgaaacgg tcacaaagcc atcttcctta ctttactcct
     1621 aaaccacctc aagatagtgc ggttatcaaa gctggatatt gtgtaaaaca aggagcagtg
     1681 atgaaaaact ggaagagaag atattttcaa ttggatgaaa acacaatagg ctacttcaaa
     1741 tctgaactgg aaaaggaacc tcttcgcgta ataccactta aagaggttca taaagtccag
     1801 gaatgtaagc aaagcgacat aatgatgagg gacaacctct ttgaaattgt aacaacgtct
     1861 cgaactttct atgtgcaggc tgatagccct gaagagatgc acagttggat taaagcagtc
     1921 tctggcgcca ttgtagcaca gcggggtccc ggcagatctg cgtcttctga gcatcccccc
     1981 ggtccttcag aatccaaaca cgctttccgt cctaccaacg cagccgccgc cacctcacat
     2041 tccacagcct ctcgcagcaa ctctttggtc tcaaccttta ccatggagaa gcgaggattt
     2101 tacgagtctc ttgccaaggt caagccaggg aacttcaagg tccagactgt ctctccaaga
     2161 gaaccagctt ccaaagtgac tgaacaagct ctgttaagac ctcaaagtaa aaatggccct
     2221 caggaaaaag attgtgacct agtagacttg gacgatgcga gccttccggt cagtgacgtg
     2281 ggtaccgcgg gcccgggatc cgccaccatg gaacaaaaac tcatctcaga agaggatctg
     2341 aatgatcagc aggacctagt ggaatgggta aatgtgttaa acaaagctat aaaaattaca
     2401 gtaccaaagc agtcagactc acagcctaat tctgataacc taagtcgcca tggtgaatgt
     2461 gggaaaaagc aagtgtctta cagaactgat attgttggtg gcgtacccat cattactccc
     2521 actcagaaag aagaagtaaa tgaatgtggt gaaagtattg acagaaataa tctgaaacgg
     2581 tcacaaagcc atcttcctta ctttactcct aaaccacctc aagatagtgc ggttatcaaa
     2641 gctggatatt gtgtaaaaca aggagcagtg atgaaaaact ggaagagaag atattttcaa
     2701 ttggatgaaa acacaatagg ctacttcaaa tctgaactgg aaaaggaacc tcttcgcgta
     2761 ataccactta aagaggttca taaagtccag gaatgtaagc aaagcgacat aatgatgagg
     2821 gacaacctct ttgaaattgt aacaacgtct cgaactttct atgtgcaggc tgatagccct
     2881 gaagagatgc acagttggat taaagcagtc tctggcgcca ttgtagcaca gcggggtccc
     2941 ggcagatctg cgtcttctga gcatcccccc ggtccttcag aatccaaaca cgctttccgt
     3001 cctaccaacg cagccgccgc cacctcacat tccacagcct ctcgcagcaa ctctttggtc
     3061 tcaaccttta ccatggagaa gcgaggattt tacgagtctc ttgccaaggt caagccaggg
     3121 aacttcaagg tccagactgt ctctccaaga gaaccagctt ccaaagtgac tgaacaagct
     3181 ctgttaagac ctcaaagtaa aaatggccct caggaaaaag attgtgacct agtagacttg
     3241 gacgatgcga gccttccggt cagtgacgtg tgaggcagaa gcgcacggag cctgcctgcc
     3301 tctgccgtcc tcagttacct ttcatgaggc ttctagccaa agatgataaa gggggaaatg
     3361 gtttttagtg cgtatattat actgcctctt aggtgtactc tttgaggatc cgccaccatg
     3421 gaacaaaaac tcatctcaga agaggatctg aatgatcagc aggacctagt ggaatgggta
     3481 aatgtgttaa acaaagctat aaaaattaca gtaccaaagc agtcagactc acagcctaat
     3541 tctgataacc taagtcgcca tggtgaatgt gggaaaaagc aagtgtctta cagaactgat
     3601 attgttggtg gcgtacccat cattactccc actcagaaag aagaagtaaa tgaatgtggt
     3661 gaaagtattg acagaaataa tctgaaacgg tcacaaagcc atcttcctta ctttactcct
     3721 aaaccacctc aagatagtgc ggttatcaaa gctggatatt gtgtaaaaca aggagcagtg
     3781 atgaaaaact ggaagagaag atattttcaa ttggatgaaa acacaatagg ctacttcaaa
     3841 tctgaactgg aaaaggaacc tcttcgcgta ataccactta aagaggttca taaagtccag
     3901 gaatgtaagc aaagcgacat aatgatgagg gacaacctct ttgaaattgt aacaacgtct
     3961 cgaactttct atgtgcaggc tgatagccct gaagagatgc acagttggat taaagcagtc
     4021 tctggcgcca ttgtagcaca gcggggtccc ggcagatctg cgtcttctga gcatcccccc
     4081 ggtccttcag aatccaaaca cgctttccgt cctaccaacg cagccgccgc cacctcacat
     4141 tccacagcct ctcgcagcaa ctctttggtc tcaaccttta ccatggagaa gcgaggattt
     4201 tacgagtctc ttgccaaggt caagccaggg aacttcaagg tccagactgt ctctccaaga
     4261 gaaccagctt ccaaagtgac tgaacaagct ctgttaagac ctcaaagtaa aaatggccct
     4321 caggaaaaag attgtgacct agtagacttg gacgatgcga gccttccggt cagtgacgtg
     4381 tgaggcagaa gcgcacggag cctgcctgcc tctgccgtcc tcagttacct ttcatgaggc
     4441 ttctagccaa agatgataaa gggggaaatg gtttttagtg cgtatattat actgcctctt
     4501 aggtgtactc tttgaggatc caccggatct agataactga tcataatcag ccataccaca
     4561 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4621 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4681 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4741 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     4801 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4861 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4921 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4981 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5041 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5101 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5161 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5221 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5281 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5341 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5401 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5461 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5521 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5641 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5701 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5761 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5821 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5881 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5941 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6001 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6061 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6121 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6181 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6241 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     6301 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6361 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6421 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6481 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6541 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6601 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6661 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6721 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6781 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6841 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6901 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6961 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     7021 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7081 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7141 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7201 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7261 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7321 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7381 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7441 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7501 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7561 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7621 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7681 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7741 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7801 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
