LOCUS       sequence-399175-        6357 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 6357)
  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 6357)
  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-PIP4K2A
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             71..865
                     /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             71..865
                     /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(125..144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     735..754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1053..1072)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1097..1144
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1473..2061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1962..1981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2236..2539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2540..2743
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2694..2714
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2782..2791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2788..3495
                     /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(2933..2951)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3201..3220
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3556..3876
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    5320..5441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5357..5376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5411..5430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5448..5903)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5585..5606)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5797..5816
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5930..6034
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6036..6357,1..36)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6036..6227
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6059..6079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6244..6357,1..22)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
       61 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
      121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
      181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
      241 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
      301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
      361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
      421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
      481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
      541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
      601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
      661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
      721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
      781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
      841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
      901 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
      961 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     1021 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     1081 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     1141 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     1201 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     1261 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     1321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2101 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2161 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     2221 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     2281 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2341 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2401 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2461 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2521 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2581 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2641 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2701 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     2761 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
     2821 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2881 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2941 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     3001 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     3061 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     3121 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     3181 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     3241 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     3301 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     3361 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     3421 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     3481 gacgagctgt acaagtccgg actcagatct cgaagcgcgg ccgcgggagc aggaggagca
     3541 gctcgagcgg cgctgggagt gcaggtggaa accatctccc caggagacgg gcgcaccttc
     3601 cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa
     3661 tttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa gcaggaggtg
     3721 atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc caaactgact
     3781 atatctccag attatgccta tggtgccact gggcacccag gcatcatccc accacatgcc
     3841 actctcgtct tcgatgtgga gcttctaaaa ctggaaagtg ctggtggtag tgctggtggt
     3901 agtgctggtg gtagtgctgg tggtagtgct ggtggtccgc gagctcaagc ttcgaattct
     3961 gcagtcgaca tggcgacccc cggcaaccta gggtcctccg tcctggcgag caagaccaag
     4021 accaagaaga agcacttcgt agcgcagaaa gtgaagctgt ttcgggccag cgacccgctg
     4081 ctcagcgtcc tcatgtgggg ggtaaaccac tcgatcaatg aactgagcca tgttcaaatc
     4141 cctgttatgt tgatgccaga tgacttcaaa gcctattcaa aaataaaggt ggacaatcac
     4201 ctttttaaca aagaaaacat gccgagccat ttcaagttta aggaatactg cccgatggtc
     4261 ttccgtaacc tgcgggagag gtttggaatt gatgatcaag atttccagaa ttccctgacc
     4321 aggagcgcac ccctccccaa cgactcccag gcccgcagtg gagctcgttt tcacacttcc
     4381 tacgacaaaa gatacatcat caagactatt accagtgaag acgtggccga aatgcacaac
     4441 atcctgaaga aataccacca gtacatagtg gaatgtcatg ggatcaccct tcttccccag
     4501 ttcttgggca tgtaccggct taatgttgat ggagttgaaa tatatgtgat agttacaaga
     4561 aatgtattca gccaccgttt gtctgtgtat aggaaatacg acttaaaggg ctctacagtg
     4621 gctagagaag ctagtgacaa agaaaaggcc aaagaactgc caactctgaa agataatgat
     4681 ttcattaatg agggccaaaa gatttatatt gatgacaaca gcaagaaggt cttcctggaa
     4741 aaactaaaaa aggatgttga gtttctggcc cagctgaagc tcatggacta cagtctgctg
     4801 gtgggaattc atgatgtgga gagagccgaa caggaggaag tggagtgtga ggagaacgat
     4861 ggggaggagg agggcgagag cgatggcacc cacccggtgg gaaccccccc agatagcccc
     4921 gggaatacac tgaacagctc accacccctg gctcccgggg agttcgatcc gaacatcgac
     4981 gtctatggaa ttaagtgcca tgaaaactcg cctaggaagg aggtgtactt catggcaatt
     5041 attgacatcc ttactcatta tgatgcaaaa aagaaagctg cccatgctgc aaaaactgtt
     5101 aaacatggcg ctggcgcgga gatctccacc gtgaacccag aacagtattc aaagcgcttt
     5161 ttggacttta ttggccacat cttgacgtaa ggatccaccg gatctagata actgatcata
     5221 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     5281 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     5341 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5401 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     5461 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5521 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5581 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5641 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5701 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5761 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5821 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5881 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5941 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6001 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     6061 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6301 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtag
//
