LOCUS       sequence-399279-        6387 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 6387)
  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 6387)
  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: mCherry-FKBP-PIP4K2BD278K
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             85..879
                     /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             85..879
                     /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(139..158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     749..768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1067..1086)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1111..1158
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1487..2075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1976..1995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2250..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2554..2757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2708..2728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2796..2805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2802..3509
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2947..2965)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3215..3234
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3570..3890
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    5364..5485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5401..5420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5455..5474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5492..5947)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5629..5650)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5841..5860
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5974..6078
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6080..6387,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6080..6271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6103..6123)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6288..6387,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
       61 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      121 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      181 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      241 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
      301 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      361 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      421 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      481 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      541 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      601 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      661 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
      721 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
      781 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
      841 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
      901 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
      961 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     1021 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     1081 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     1141 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     1201 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     1261 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     1321 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1621 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2161 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2761 ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc aagggcgagg
     2821 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     2881 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2941 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     3001 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     3061 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     3121 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     3181 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     3241 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     3301 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     3361 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     3421 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     3481 ccaccggcgg catggacgag ctgtacaagt ccggactcag atctcgaagc gcggccgcgg
     3541 gagcaggagg agcagctcga gcggcgctgg gagtgcaggt ggaaaccatc tccccaggag
     3601 acgggcgcac cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg
     3661 aagatggaaa gaaatttgat tcctcccggg acagaaacaa gccctttaag tttatgctag
     3721 gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga
     3781 gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca
     3841 tcccaccaca tgccactctc gtcttcgatg tggagcttct aaaactggaa agtgctggtg
     3901 gtagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggt ccgcgagctc
     3961 aagcttcgaa ttctgcagtc gacatgtcgt ccaactgcac cagcaccacg gcggtggcgg
     4021 tggcgccgct cagcgccagc aagaccaaga ccaagaagaa gcatttcgtg tgccagaaag
     4081 tgaagctatt ccgggccagc gagccgatcc tcagcgtcct gatgtggggg gtgaaccaca
     4141 cgatcaatga gctgagcaat gttcctgttc ctgtcatgct aatgccagat gacttcaaag
     4201 cctacagcaa gatcaaggtg gacaatcatc tcttcaataa ggagaacctg cccagccgct
     4261 ttaagtttaa ggagtattgc cccatggtgt tccgaaacct tcgggagagg tttggaattg
     4321 atgatcagga ttaccagaat tcagtgacgc gcagcgcccc catcaacagt gacagccagg
     4381 gtcggtgtgg cacgcgtttc ctcaccacct acgaccggcg ctttgtcatc aagactgtgt
     4441 ccagcgagga cgtggcggag atgcacaaca tcttaaagaa ataccaccag tttatagtgg
     4501 agtgtcatgg caacacgctt ttgccacagt tcctgggcat gtaccgcctg accgtggatg
     4561 gtgtggaaac ctacatggtg gttaccagga acgtgttcag ccatcggctc actgtgcatc
     4621 gcaagtatga cctcaagggt tctacggttg ccagagaagc gagcgacaag gagaaggcca
     4681 aggacttgcc aacattcaaa gacaatgact tcctcaatga agggcagaag ctgcatgtgg
     4741 gagaggagag taaaaagaac ttcctggaga aactgaagcg ggacgttgag ttcttggcac
     4801 agctgaagat catgaaatac agcctgctgg tgggcatcca cgacgtggac cgggcagagc
     4861 aggaggagat ggaggtggag gagcgggcag aggacgagga gtgtgagaat gatggggtgg
     4921 gtggcaacct actctgctcc tatggcacac ctccggacag ccctggcaac ctcctcagct
     4981 ttcctcggtt ctttggtcct ggggaattcg acccctctgt tgacgtctat gccatgaaaa
     5041 gccatgaaag ttcccccaag aaggaggtgt atttcatggc catcattgat atcctcacgc
     5101 catacgatac aaagaagaaa gctgcacatg ctgccaaaac ggtgaaacac ggggcagggg
     5161 ccgagatctc gactgtgaac cctgagcagt actccaaacg cttcaacgag tttatgtcca
     5221 acatcctgac gtagggatcc accggatcta gataactgat cataatcagc cataccacat
     5281 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5341 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5401 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5461 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     5521 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5581 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5641 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5701 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5761 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5821 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5881 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5941 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6001 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6061 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6121 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6181 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6241 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6301 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6361 aggccgaggc cgcctcggcc tctgagc
//
