LOCUS       sequence-398134-        6261 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 6261)
  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 6261)
  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-Kina
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..311
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     268..288
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(313..417)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      444..899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(531..550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     741..762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    906..1027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(917..936)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     971..990
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2375..2695)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(2756..3463)
                     /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(3031..3050)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3300..3318
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3460..3469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3508..3711)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3537..3557)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3712..4015
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5107..5154
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5179..5198
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5386..6180)
                     /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             complement(5386..6180)
                     /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(5497..5516)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6107..6126
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6215..6261,1..311))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6229..6261,1..103)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
       61 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
      121 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
      241 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agctggttct
      301 ttccgcctca ggactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      361 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      421 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
      481 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      541 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      601 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      661 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      721 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
      781 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
      841 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
      901 cgcgttaaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      961 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1021 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1081 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagttatc
     1141 tagatccggt ggatccttac gtcaagatgt ggccaataaa gtccaaaaag cgctttgaat
     1201 actgttctgg gttcacggtg gagatctccg cgccagcgcc atgtttaaca gtttttgcag
     1261 catgggcagc tttctttttt gcatcataat gagtaaggat gtcaataatt gccatgaagt
     1321 acacctcctt cctaggcgag ttttcatggc acttaattcc atagacgtcg atgttcggat
     1381 cgaactcccc gggagccagg ggtggtgagc tgttcagtgt attcccgggg ctatctgggg
     1441 gggttcccac cgggtgggtg ccatcgctct cgccctcctc ctccccatcg ttctcctcac
     1501 actccacttc ctcctgttcg gctctctcca catcatgaat tcccaccagc agactgtagt
     1561 ccatgagctt cagctgggcc agaaactcaa catccttttt tagtttttcc aggaagacct
     1621 tcttgctgtt gtcatcaata taaatctttt ggccctcatt aatgaaatca ttatctttca
     1681 gagttggcag ttctttggcc ttttctttgt cactagcttc tctagccact gtagagccct
     1741 ttaagtcgta tttcctatac acagacaaac ggtggctgaa tacatttctt gtaactatca
     1801 catatatttc aactccatca acattaagcc ggtacatgcc caagaactgg ggaagaaggg
     1861 tgatcccatg acattccact atgtactggt ggtatttctt caggatgttg tgcatttcgg
     1921 ccacgtcttc actggtaata gtcttgatga tgtatctttt gtcgtaggaa gtgtgaaaac
     1981 gagctccact gcgggcctgg gagtcgttgg ggaggggtgc gctcctggtc agggaattct
     2041 ggaaatcttg atcatcaatt ccaaacctct cccgcaggtt acggaagacc atcgggcagt
     2101 attccttaaa cttgaaatgg ctcggcatgt tttctttgtt aaaaaggtga ttgtccacct
     2161 ttatttttga ataggctttg aagtcatctg gcatcaacat aacagggatt tgaacatggc
     2221 tcagttcatt gatcgagtgg tttacccccc acatgaggac gctgagcagc gggtcgctgg
     2281 cgtcgactgc agaattcgaa gcttgagctc gcggaccacc agcactacca ccagcactac
     2341 caccagcact accaccagca ctaccaccag cactttccag ttttagaagc tccacatcga
     2401 agacgagagt ggcatgtggt gggatgatgc ctgggtgccc agtggcacca taggcataat
     2461 ctggagatat agtcagtttg gctctctgac ccacactcat ctgggcaacc ccttcttccc
     2521 agcctcggat cacctcctgc ttgcctagca taaacttaaa gggcttgttt ctgtcccggg
     2581 aggaatcaaa tttctttcca tcttcaagca tcccggtgta gtgcaccacg caggtctggc
     2641 cgcgcttggg gaaggtgcgc ccgtctcctg gggagatggt ttccacctgc actcccagcg
     2701 ccgctcgagc tgctcctcct gctcccgcgg ccgcgcttcg agatctgagt ccggacttgt
     2761 acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga
     2821 tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg taggcgccgg
     2881 gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc
     2941 cgtccttcag cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca
     3001 tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg
     3061 ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc agggaggagt
     3121 cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc
     3181 cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct
     3241 tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg gggccaccct
     3301 tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct
     3361 cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg caccttgaag cgcatgaact
     3421 ccttgatgat ggccatgtta tcctcctcgc ccttgctcac catggtggcg accggtagcg
     3481 ctagcggatc tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg
     3541 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     3601 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     3661 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     3721 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     3781 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     3841 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     3901 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     3961 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     4021 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca
     4081 tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     4981 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     5041 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     5101 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     5161 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     5221 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     5281 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     5341 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     5401 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     5461 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     5521 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     5581 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     5641 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     5701 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     5761 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     5821 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     5881 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     5941 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     6001 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     6061 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     6121 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     6181 gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa
     6241 agcctcctca ctacttctgg a
//
