LOCUS       sequence-318718-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector, contains two tandem repeats of OSH2 PH 
            domain (PI(4)P biosensor). Fused to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  AUTHORS   Balla A, Kim YJ, Varnai P, Szentpetery Z, Knight Z, Shokat KM, Balla
            T
  TITLE     Maintenance of hormone-sensitive phosphoinositide pools in the 
            plasma membrane requires phosphatidylinositol 4-kinase IIIalpha.
  JOURNAL   Mol Biol Cell. 2008 Feb;19(2):711-21. doi: 10.1091/mbc.e07-07-0713. 
            Epub 2007 Dec 12.
  PUBMED    18077555
REFERENCE   2  (bases 1 to 5739)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1091/mbc.e07-07-0713"; journalName: "Mol Biol Cell"; date: 
            "2008-02"; volume: "19"; issue: "2"; pages: "711-21"
COMMENT     Sequence Label: GFP-OSH2-2xPH
FEATURES             Location/Qualifiers
     source          1..5739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        111..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      657..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(663..665,666..668,669..1379)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(663..665,666..668,669..1379)
                     /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(708..729)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(969..988)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1316..1337
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2577..2698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2614..2633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2668..2687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2705..3160)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2842..2863)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3054..3073
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3187..3291
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3293..3650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3293..3484
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3316..3336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3501..3636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3685..4479
                     /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             3685..4479
                     /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(3739..3758)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4349..4368
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4667..4686)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4711..4758
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5087..5675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5576..5595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca
      661 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      721 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      781 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      841 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      901 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      961 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1021 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1081 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1141 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1201 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1261 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1321 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1381 ccggactcag atctcgagaa ccgtcgaaca acgttacacc ggaaattaag ttaaagaatc
     1441 tacttgaaaa gaatctgaga gaacaagcta ttgttcacga agatgtcgca tcaagtaagc
     1501 ctcctacgta taaggggttt ttaaagaaat ggacaaattt tgcccatggc tacaaattac
     1561 gttggttcat tttaagtggc gacggaaatc tgtcatatta caaagaccaa tcgcatgttg
     1621 ataggcctcg tggtacttta aaagtttcga cgtgccgctt acacatcgat tcttcagaaa
     1681 aactgaattt cgagttgcta ggtggcatca caggcacaac aaggtggcgc ttaaagggca
     1741 atcatcccat tgaaacgact aggtgggtca atgctataca aagcgctatt agattcgcca
     1801 aagacaagga gatattaaac aaaaaaaagg cagtaccacc atcattggcc ttgaagaaca
     1861 aatctccagc cctgatatcg cattccaaaa cacaaggatc tcttcctgtg aattccaagc
     1921 taccgtcgaa caacgttaca ccggaaatta agttaaagaa tctacttgaa aagaatctga
     1981 gagaacaagc tattgttcac gaagatgtcg catcaagtaa gcctcctacg tataaggggt
     2041 ttttaaagaa atggacaaat tttgcccatg gctacaaatt acgttggttc attttaagtg
     2101 ccgacggaaa tctgtcatat tacaaagacc aatcgcatgt tgataggcct cgtggtactt
     2161 taaaagtttc gacgtgccgt ttacacatcg attcttcaga aaaactgaat ttcgagttac
     2221 taggtggcat cacaggcaca acaaggtggc gcttgaaggg caatcatccc attgaaacga
     2281 ctaggtgggt caatgctata caaagcgcta ttagattcgc caaagacaag gagatattaa
     2341 acaaaaaaaa ggcagtacca ccatcattgg ccttgaagaa caaatctcca gccctgatat
     2401 cgcattccaa aacacaagga tctcttcctt aaggtaccgc gggcccggga tccaccggat
     2461 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2521 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     2701 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2761 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2821 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2881 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2941 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3001 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3061 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3121 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3301 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3481 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3541 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3601 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3661 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3721 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3781 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3841 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3901 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3961 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4021 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4081 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4141 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4201 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4261 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4321 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4381 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4441 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4501 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4561 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4621 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4681 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4741 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4801 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4861 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4921 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4981 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5701 ggttcctggc cttttgctgg ccttttgctc acatgttct
//
