LOCUS       sequence-360288-        6360 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Recruitable human type IV 5-phosphatase enzyme (catalytically 
            inactive).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  AUTHORS   Walpole GFW, Pacheco J, Chauhan N, Clark J, Anderson KE, Abbas YM, 
            Brabant-Kirwan D, Montano-Rendon F, Liu Z, Zhu H, Brumell JH, 
            Deiters A, Stephens LR, Hawkins PT, Hammond GRV, Grinstein S, Fairn 
            GD
  TITLE     Kinase-independent synthesis of 3-phosphorylated phosphoinositides 
            by a phosphotransferase.
  JOURNAL   Nat Cell Biol. 2022 Apr 28. pii: 10.1038/s41556-022-00895-y. doi: 
            10.1038/s41556-022-00895-y.
  PUBMED    35484249
REFERENCE   2  (bases 1 to 6360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2022 Apr 28. pii: 10.1038/s41556-022-00895-y. doi: 
            10.1038/s41556-022-00895-y."
COMMENT     Sequence Label: mRFP-FKBP-INPP5E(D556A)
FEATURES             Location/Qualifiers
     source          1..6360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             618..1292
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             618..1292
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(748..766)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1016..1035
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1210..1233
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             join(1350..1352,1353..1673)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    3153..3274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3190..3209)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3244..3263
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3281..3736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3418..3439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3630..3649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3763..3867
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3869..4226
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3869..4060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3892..3912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4077..4212
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4261..5055
                     /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             4261..5055
                     /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(4315..4334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4925..4944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5243..5262)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5287..5334
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5663..6251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6152..6171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 cgctaccggt cgccaccatg gcctcctccg aggacgtcat caaggagttc atgcgcttca
      661 aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
      781 ccttcgcctg ggacatcctg tcccctcagt tccagtacgg ctccaaggcc tacgtgaagc
      841 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      901 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
      961 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1021 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcggatgtac cccgaggacg
     1081 gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc cactacgacg
     1141 ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc gcctacaaga
     1201 ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1261 agcgcgccga gggccgccac tccaccggcg cctccggact cagatctcga agcgcggccg
     1321 cgggagcagg aggagcagct cgagcggcga tgggagtgca ggtggaaacc atctccccag
     1381 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     1441 ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt aagtttatgc
     1501 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     1561 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     1621 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaagctcgag
     1681 gcgcagcagc gggagcagga ggagcaggtc gatcggatct tgcagactac aagctccgcg
     1741 cgcagccgct cctggtgcgg gcccacagca gcctgggccc cggccggccg cggagccccc
     1801 tggcctgcga cgactgttcc cttcgctcgg ccaaatcctc cttcagcctc ctggcgccca
     1861 tccgcagcaa ggacgtccgc agcaggagtt acctggaggg cagcctcctg gccagcgggg
     1921 ccctgttggg ggcggatgag ctggcccgct acttcccaga ccggaacgtg gcactcttcg
     1981 tggccacctg gaacatgcag ggccagaagg agctcccgcc cagcctggac gagttcctgc
     2041 tcccagccga ggccgactat gcccaggacc tgtatgtcat cggggtccag gagggctgtt
     2101 ctgacaggcg ggagtgggag actcgtctgc aggagacgct gggcccgcac tatgtgctgc
     2161 tgtcctcggc ggcccacggc gtgctctaca tgtcgctctt catccgcagg gacctcatct
     2221 ggttctgctc agaggtggag tgctccacgg tgaccacacg catcgtgtct cagatcaaga
     2281 ccaagggggc cttgggcatc agcttcacct tttttggcac ttccttcctc ttcatcacgt
     2341 cccacttcac ctcaggtgac gggaaggtgg cggagcggct gctggactac accaggactg
     2401 tacaagccct ggtcctgccc agaaatgtgc ccgacaccaa cccctatcgc tccagcgcag
     2461 cggacgtcac cacccgcttc gatgaggtgt tctggtttgg agacttcaac ttccgcctga
     2521 gtggcgggcg cacagtcgtg gacgccctcc tgtgccaggg cctggtggtg gacgtgccgg
     2581 cgctgctgca gcacgaccag ctcatccggg agatgcggaa agggtccatc ttcaagggct
     2641 tccaggagcc ggacatccac ttcctcccat catacaagtt tgacatcggg aaggacacgt
     2701 acgacagcac ctccaagcag aggacgccct catacacggc ccgcgtcttg tacagaagcc
     2761 gccacaaggg tgacatctgt cctgtgagct actcttcctg ccccgggatc aagacgtccg
     2821 accaccgccc tgtgtatggc ctcttccggg tgaaagtgag gccggggcga gacaacattc
     2881 cgttggcagc tggcaaattt gatagagaac tgtacttact aggaattaaa agacggattt
     2941 cgaaggagat tcagaggcag caagcactac agagtcagaa ctccagcacc atcgcctccg
     3001 tttcttgagg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     3061 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3121 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3241 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3301 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3361 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3421 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3481 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3541 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3601 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3661 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3721 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3781 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3841 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4081 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4141 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4201 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4261 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4321 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4381 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4441 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4501 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4561 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4621 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4681 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4741 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4801 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4861 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4921 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4981 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5041 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5101 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5161 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5221 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5281 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5341 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5401 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5461 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6301 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
//
