LOCUS       sequence-399137-        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-PIP4K2B
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             137..931
                     /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             137..931
                     /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(191..210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     801..820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1119..1138)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1163..1210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1539..2127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2028..2047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2302..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2606..2809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2848..2857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2854..3561
                     /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(2999..3017)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3267..3286
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3622..3942
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    5416..5537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5453..5472)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5507..5526
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5544..5999)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5681..5702)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5893..5912
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6026..6130
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6132..6387,1..102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6132..6323
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6155..6175)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6340..6387,1..88)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
       61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
      121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
      181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
      241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
      301 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
      361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
      421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
      481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
      541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
      601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
      661 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
      721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
      781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
      841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
      901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
      961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     1021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1081 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     1141 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     1201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     1261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     1321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     1381 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2221 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2821 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggaggataac
     2881 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2941 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     3001 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     3061 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     3121 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     3181 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     3241 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     3301 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     3361 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     3421 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     3481 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     3541 ggcatggacg agctgtacaa gtccggactc agatctcgaa gcgcggccgc gggagcagga
     3601 ggagcagctc gagcggcgct gggagtgcag gtggaaacca tctccccagg agacgggcgc
     3661 accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga
     3721 aagaaatttg attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag
     3781 gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa
     3841 ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca
     3901 catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaagtgctgg tggtagtgct
     3961 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtccgcgagc tcaagcttcg
     4021 aattctgcag tcgacatgtc gtccaactgc accagcacca cggcggtggc ggtggcgccg
     4081 ctcagcgcca gcaagaccaa gaccaagaag aagcatttcg tgtgccagaa agtgaagcta
     4141 ttccgggcca gcgagccgat cctcagcgtc ctgatgtggg gggtgaacca cacgatcaat
     4201 gagctgagca atgttcctgt tcctgtcatg ctaatgccag atgacttcaa agcctacagc
     4261 aagatcaagg tggacaatca tctcttcaat aaggagaacc tgcccagccg ctttaagttt
     4321 aaggagtatt gccccatggt gttccgaaac cttcgggaga ggtttggaat tgatgatcag
     4381 gattaccaga attcagtgac gcgcagcgcc cccatcaaca gtgacagcca gggtcggtgt
     4441 ggcacgcgtt tcctcaccac ctacgaccgg cgctttgtca tcaagactgt gtccagcgag
     4501 gacgtggcgg agatgcacaa catcttaaag aaataccacc agtttatagt ggagtgtcat
     4561 ggcaacacgc ttttgccaca gttcctgggc atgtaccgcc tgaccgtgga tggtgtggaa
     4621 acctacatgg tggttaccag gaacgtgttc agccatcggc tcactgtgca tcgcaagtat
     4681 gacctcaagg gttctacggt tgccagagaa gcgagcgaca aggagaaggc caaggacttg
     4741 ccaacattca aagacaatga cttcctcaat gaagggcaga agctgcatgt gggagaggag
     4801 agtaaaaaga acttcctgga gaaactgaag cgggacgttg agttcttggc acagctgaag
     4861 atcatggact acagcctgct ggtgggcatc cacgacgtgg accgggcaga gcaggaggag
     4921 atggaggtgg aggagcgggc agaggacgag gagtgtgaga atgatggggt gggtggcaac
     4981 ctactctgct cctatggcac acctccggac agccctggca acctcctcag ctttcctcgg
     5041 ttctttggtc ctggggaatt cgacccctct gttgacgtct atgccatgaa aagccatgaa
     5101 agttccccca agaaggaggt gtatttcatg gccatcattg atatcctcac gccatacgat
     5161 acaaagaaga aagctgcaca tgctgccaaa acggtgaaac acggggcagg ggccgagatc
     5221 tcgactgtga accctgagca gtactccaaa cgcttcaacg agtttatgtc caacatcctg
     5281 acgtagggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     5341 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5401 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5461 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5521 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     5581 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5641 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5701 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5761 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5821 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5881 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5941 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6121 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     6181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6241 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6301 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6361 agttccgccc attctccgcc ccatggc
//
