LOCUS       sequence-269437-        5709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal half of split PI-PLC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5709)
  AUTHORS   Zewe JP, Miller AM, Sangappa S, Wills RC, Goulden BD, Hammond GRV
  TITLE     Probing the subcellular distribution of phosphatidylinositol reveals
            a surprising lack at the plasma membrane.
  JOURNAL   J Cell Biol. 2020 Mar 2;219(3). pii: 133808. doi: 
            10.1083/jcb.201906127.
  PUBMED    32211893
REFERENCE   2  (bases 1 to 5709)
  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"; journalName: "J Cell 
            Biol."; date: "2020-03-2"; volume: "219(3). pii: 133808. doi"; 
            pages: " 10.1083/jcb.201906127"
COMMENT     Sequence Label: TagBFP2-C1-FKB-PI-PLCN187
FEATURES             Location/Qualifiers
     source          1..5709
                     /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             613..1314
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             1375..1695
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    2497..2618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2588..2607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2625..3080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2762..2783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2974..2993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3107..3211
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3213..3570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3213..3404
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3236..3256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3421..3556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3605..4399
                     /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             3605..4399
                     /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(3659..3678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4269..4288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4587..4606)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4631..4678
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5007..5595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5496..5515
                     /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 cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
      661 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
      721 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
      781 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
      841 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
      901 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      961 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     1021 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     1081 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     1141 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     1201 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     1261 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattccgga
     1321 ctcagatctc gaagcgcggc cgcgggagca ggaggagcag ctcgagcggc gctgggagtg
     1381 caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc
     1441 gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga
     1501 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     1561 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     1621 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     1681 cttctaaaac tggaaagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     1741 ggtagtgctg gtggtccgcg agctcaagct tcgtcgctta ataactggaa taagccaata
     1801 aagaactctg taactacaaa acaatggatg tccgctctac ctgacacaac aaacctagca
     1861 gcgctctcta taccaggtac acatgatacg atgagctata acggagacat aacatggaca
     1921 ttaaccaaac cactagctca aacacaaacg atgtcattgt accaacaact agaagcagga
     1981 atacggtaca tcgatattag agcaaaagac aatctcaaca tttaccatgg gccaattttt
     2041 ttaaatgcat cactttcagg tgtattagaa acgattactc aatttttaaa gaaaaatcca
     2101 aaagaaacca ttattatgcg tttaaaggac gagcaaaaca gcaacgatag ttttgattat
     2161 cggatccaac cactaatcaa catttataaa gattattttt acactactcc cagaactgac
     2221 acgagcaata aaatccctac attaaaagat gtccgcggaa aaatattatt actttcagag
     2281 aaccacacaa aaaagccatt agtcattaac tcacgcaaat tcggcatgca gttctagaat
     2341 tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
     2401 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2461 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2521 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2581 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2641 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2701 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2761 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2821 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2881 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2941 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3001 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3061 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3181 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3241 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3421 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3481 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3541 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3601 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3661 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3721 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3781 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3841 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3901 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3961 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4021 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4081 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4141 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4201 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4261 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4321 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4381 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4441 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4501 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4561 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4621 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4681 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4741 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4801 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4861 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4921 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4981 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5041 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5101 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5161 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5221 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5281 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5341 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5401 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5461 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5521 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5581 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5641 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5701 gccatgcat
//
