LOCUS       sequence-399530-        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-HD
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..873
                     /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             79..873
                     /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(133..152)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     743..762
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1061..1080)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1105..1152
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1481..2069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1970..1989
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2244..2547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2548..2751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2702..2722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2790..2799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2796..3497
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             3558..3878
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    5223..5344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5260..5279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5314..5333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5351..5806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5488..5509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5700..5719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5833..5937
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5939..6252,1..44)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5939..6130
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5962..5982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6147..6252,1..30)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
       61 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      121 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      181 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      241 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      301 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      361 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      421 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      481 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      541 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      601 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
      661 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
      721 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
      781 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
      841 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
      901 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
      961 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1021 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1081 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1141 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1201 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1261 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1321 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1381 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1441 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1501 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1561 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1621 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1681 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1741 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1801 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1861 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1921 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1981 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2041 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2101 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2161 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2221 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2281 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2341 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2401 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2461 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2521 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2581 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2641 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2701 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     2761 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcgagctg attaaggaga
     2821 acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat
     2881 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
     2941 gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga
     3001 ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct
     3061 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     3121 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat
     3181 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt
     3241 accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga
     3301 gccatctgat cgcaaacgcc aagaccacat atagatccaa gaaacccgct aagaacctca
     3361 agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg
     3421 agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac
     3481 tggggcacaa gcttaattcc ggactcagat ctcgaagcgc ggccgcggga gcaggaggag
     3541 cagctcgagc ggcgctggga gtgcaggtgg aaaccatctc cccaggagac gggcgcacct
     3601 tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga
     3661 aatttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg
     3721 tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga
     3781 ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg
     3841 ccactctcgt cttcgatgtg gagcttctaa aactggaaag tgctggtggt agtgctggtg
     3901 gtagtgctgg tggtagtgct ggtggtagtg ctggtggtcc gcgagctcaa gcttcgaatt
     3961 ctgcagtcga cggtaccgcg ggcccgggat ccctgaaggg tgccatccag ctgggcatcg
     4021 gctacaccgt gggccacctg agctccaagc ccgaacgccg cgtgctcatg caggacttct
     4081 acgtggtgga gagcatcttc ttccccagcg aaggcagcaa cctcaccccc gcccaccact
     4141 tccaggactt ccgcttcaag acctatgcac ctgtcgcctt ccgctacttc cgggagctct
     4201 ttgggatccg gccagatgat tacttgtact ccctgtgcaa tgagccgctg atcgagctgt
     4261 ccaacccggg cgccagtggc tccctcttct acgtcaccag cgacgacgag ttcatcatca
     4321 agaccgtcat gcacaaggag gccgagttcc tgcagaagct gctccctggc tactacatga
     4381 acctcaacca gaacccgcgg acgctgctgc ccaagttcta tgggctgtac tgcgtgcagt
     4441 cggggggcaa gaacatccgc gtcgtggtca tgaacaacat cctgccccgc gtggtcaaga
     4501 tgcacctcaa gttcgacctc aagggctcca cctacaagcg gcgcgccagc aagaaggaga
     4561 aggagaagag cttccccacc tacaaggacc tggacttcat gcaggacatg cccgaggggc
     4621 tcctgctgga cgccgacacc ttcagcgccc tggtcaagac gctgcaggac gactgcctgg
     4681 tcctggaaag tttcaagatc atggactaca gcctgctgct gggcgtgcac aacatcgacc
     4741 agcacgagcg cgagcggcag gcgcagggcg cccagagcac ctcagatgag aagcggcctg
     4801 tgggccagaa ggcgctctac tccacggcca tggagtccat ccagggtggc gccgcgcgcg
     4861 gggaggccat cgaatcggat gacacgatgg gcgggatccc cgctgtgaac ggccgcgggg
     4921 agcggctgct gctgcacatt ggcatcatcg acatcctgca gtcctacagg ttcatcaaga
     4981 aactggagca cacctggaag gccctcgtcc acgatgggga cacggtgtcc gtccaccgcc
     5041 ccagcttcta tgccgagcgc tttttcaagt tcatgagcaa cacggtcttt taaggatcca
     5101 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     5161 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     5221 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5281 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5341 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5401 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5461 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5521 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5581 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5641 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     5701 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     5761 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     5821 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5881 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     5941 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     6001 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     6061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6121 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     6181 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6241 ctgagctatt cc
//
