LOCUS       sequence-399168-        6252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses recruitable fluorescent protein-tagged enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6252)
  AUTHORS   Wills RC, Doyle CP, Zewe JP, Pacheco J, Hansen SD, Hammond GRV
  TITLE     A novel homeostatic mechanism tunes PI(4,5)P2-dependent signaling at
            the plasma membrane.
  JOURNAL   J Cell Sci. 2023 Aug 3:jcs.261494. doi: 10.1242/jcs.261494.
  PUBMED    37534432
REFERENCE   2  (bases 1 to 6252)
  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 Sci.
            2023 Aug 3:jcs.261494. doi: 10.1242/jcs.261494."
COMMENT     Sequence Label: TagBFP2-FKBP-PIP5K1C_Kina-HDD316K
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        450..653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     604..624
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             698..1399
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             1460..1780
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3125..3246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3162..3181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3216..3235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3253..3708)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3390..3411)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3602..3621
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3735..3839
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3841..4198
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3841..4032
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3864..3884)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4049..4184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4233..5027
                     /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             4233..5027
                     /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(4287..4306)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4897..4916
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5215..5234)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5259..5306
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5635..6223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6124..6143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
       61 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
      121 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      181 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      241 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      301 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      361 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      421 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      481 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      541 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      601 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      661 tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcgagc tgattaagga
      721 gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac
      781 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
      841 gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa
      901 gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg
      961 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     1021 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac
     1081 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct
     1141 gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg
     1201 gagccatctg atcgcaaacg ccaagaccac atatagatcc aagaaacccg ctaagaacct
     1261 caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa
     1321 cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa
     1381 actggggcac aagcttaatt ccggactcag atctcgaagc gcggccgcgg gagcaggagg
     1441 agcagctcga gcggcgctgg gagtgcaggt ggaaaccatc tccccaggag acgggcgcac
     1501 cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa
     1561 gaaatttgat tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga
     1621 ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact
     1681 gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca
     1741 tgccactctc gtcttcgatg tggagcttct aaaactggaa agtgctggtg gtagtgctgg
     1801 tggtagtgct ggtggtagtg ctggtggtag tgctggtggt ccgcgagctc aagcttcgaa
     1861 ttctgcagtc gacggtaccg cgggcccggg atccctgaag ggtgccatcc agctgggcat
     1921 cggctacacc gtgggccacc tgagctccaa gcccgaacgc cgcgtgctca tgcaggactt
     1981 ctacgtggtg gagagcatct tcttccccag cgaaggcagc aacctcaccc ccgcccacca
     2041 cttccaggac ttccgcttca agacctatgc acctgtcgcc ttccgctact tccgggagct
     2101 ctttgggatc cggccagatg attacttgta ctccctgtgc aatgagccgc tgatcgagct
     2161 gtccaacccg ggcgccagtg gctccctctt ctacgtcacc agcgacgacg agttcatcat
     2221 caagaccgtc atgcacaagg aggccgagtt cctgcagaag ctgctccctg gctactacat
     2281 gaacctcaac cagaacccgc ggacgctgct gcccaagttc tatgggctgt actgcgtgca
     2341 gtcggggggc aagaacatcc gcgtcgtggt catgaacaac atcctgcccc gcgtggtcaa
     2401 gatgcacctc aagttcgacc tcaagggctc cacctacaag cggcgcgcca gcaagaagga
     2461 gaaggagaag agcttcccca cctacaagga cctggacttc atgcaggaca tgcccgaggg
     2521 gctcctgctg gacgccgaca ccttcagcgc cctggtcaag acgctgcagg acgactgcct
     2581 ggtcctggaa agtttcaaga tcatgaaata cagcctgctg ctgggcgtgc acaacatcga
     2641 ccagcacgag cgcgagcggc aggcgcaggg cgcccagagc acctcagatg agaagcggcc
     2701 tgtgggccag aaggcgctct actccacggc catggagtcc atccagggtg gcgccgcgcg
     2761 cggggaggcc atcgaatcgg atgacacgat gggcgggatc cccgctgtga acggccgcgg
     2821 ggagcggctg ctgctgcaca ttggcatcat cgacatcctg cagtcctaca ggttcatcaa
     2881 gaaactggag cacacctgga aggccctcgt ccacgatggg gacacggtgt ccgtccaccg
     2941 ccccagcttc tatgccgagc gctttttcaa gttcatgagc aacacggtct tttaaggatc
     3001 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3061 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3121 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3181 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3241 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3301 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3361 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3421 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3481 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3541 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3601 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3661 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3721 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3781 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3841 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3901 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3961 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4021 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4081 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4141 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4201 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4261 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4321 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4381 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4441 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4501 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4561 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4621 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4681 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4741 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4801 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4861 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4921 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4981 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5041 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5101 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5161 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5221 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5281 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5341 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5401 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5461 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5761 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6241 ctttttacgg tt
//
