LOCUS       sequence-399521-        6413 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 6413)
  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 6413)
  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-PIP4K2C
FEATURES             Location/Qualifiers
     source          1..6413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             154..948
                     /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             154..948
                     /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(208..227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     818..837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1136..1155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1180..1227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1556..2144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2045..2064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2319..2622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2623..2826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2777..2797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2865..2874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2871..3578
                     /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(3016..3034)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3284..3303
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3639..3959
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    5459..5580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5496..5515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5550..5569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5587..6042)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5724..5745)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5936..5955
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6069..6173
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6175..6413,1..119)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6175..6366
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6198..6218)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6383..6413,1..105)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
       61 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
      121 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      181 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      241 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      301 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
      361 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      421 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      481 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      541 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      601 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
      661 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
      721 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
      781 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
      841 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
      901 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
      961 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1021 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     1081 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     1141 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1201 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     1261 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     1321 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     1381 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     1441 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1681 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1981 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2041 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2101 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2161 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2221 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     2281 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2341 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2401 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2461 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2521 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2581 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2641 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2701 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2761 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2821 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca
     2881 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     2941 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3001 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     3061 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     3121 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     3181 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     3241 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     3301 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     3361 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     3421 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     3481 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     3541 gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga tctcgaagcg
     3601 cggccgcggg agcaggagga gcagctcgag cggcgctggg agtgcaggtg gaaaccatct
     3661 ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg cactacaccg
     3721 ggatgcttga agatggaaag aaatttgatt cctcccggga cagaaacaag ccctttaagt
     3781 ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc cagatgagtg
     3841 tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc actgggcacc
     3901 caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta aaactggaaa
     3961 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtc
     4021 cgcgagctca agcttcgaat tctgcagtcg acggtaccgc gatggcgtcc tcctcggtcc
     4081 caccagccac ggtatcggcg gcgacagcag gccccggccc aggtttcggc ttcgcctcca
     4141 agaccaagaa gaagcatttc gtgcagcaga aggtgaaggt gttccgggcg gccgacccgc
     4201 tggtgggtgt gttcctgtgg ggcgtagccc actcgatcaa tgagctcagc caggtgcctc
     4261 ccccggtgat gctgctgcca gatgacttta aggccagctc caagatcaag gtcaacaatc
     4321 accttttcca cagggaaaat ctgcccagtc atttcaagtt caaggagtat tgtccccagg
     4381 tcttcaggaa cctccgtgat cgatttggca ttgatgacca agattacttg gtgtccctta
     4441 cccgaaaccc ccccagcgaa agtgaaggca gtgatggtcg cttccttatc tcctacgatc
     4501 ggactctggt catcaaagaa gtatccagtg aggacattgc tgacatgcat agcaacctct
     4561 ccaactatca ccagtacatt gtgaagtgcc atggcaacac gcttctgccc cagttcctgg
     4621 ggatgtaccg agtcagtgtg gacaacgaag acagctacat gcttgtgatg cgcaatatgt
     4681 ttagccaccg tcttcctgtg cacaggaagt atgacctcaa gggttcccta gtgtcccggg
     4741 aagccagcga taaggaaaag gttaaagaat tgcccaccct taaggatatg gactttctca
     4801 acaagaacca gaaagtatat attggtgaag aggagaagaa aatatttctg gagaagctga
     4861 agagagatgt ggagtttcta gtgcagctga agatcatgga ctacagcctt ctgctaggca
     4921 tccacgacat cattcggggc tctgaaccag aggaggaagc gcccgtgcgg gaggatgagt
     4981 cagaggtgga tggggactgc agcctgactg gacctcctgc tctggtgggc tcctatggca
     5041 cctccccaga gggtatcgga ggctacatcc attcccatcg gcccctgggc ccaggagagt
     5101 ttgagtcctt cattgatgtc tatgccatcc ggagtgctga aggagccccc cagaaggagg
     5161 tctacttcat gggcctcatt gatatcctta cacagtatga tgccaagaag aaagcagctc
     5221 atgcagccaa aactgtcaag catggggctg gggcagagat ctctactgtc catccggagc
     5281 agtatgctaa gcgattcctg gattttatta ccaacatctt tgcctaacgg gatccaccgg
     5341 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     5401 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     5461 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5521 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5581 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5641 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5701 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5761 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5821 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5881 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5941 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6001 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     6061 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6121 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     6181 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     6301 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     6361 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg ccc
//
