LOCUS       Exported                7065 bp ds-DNA     circular SYN 18-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Hammond Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 16, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7065
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..589
                     /label=CMV
                     /note="CMV"
     primer_bind     519..539
                     /label=CMV-F
     primer_bind     complement(597..636)
                     /label=Cherry delrev 1
     CDS             616..1314
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     616..639
                     /label=TagBFP fwd 1
     primer_bind     616..639
                     /label=Fragment.FOR
     primer_bind     complement(715..735)
                     /label=BFP rev seq 1
     primer_bind     1203..1225
                     /label=BFP fwd seq 1
     primer_bind     complement(1292..1314)
                     /label=TagBFP rev 1
     primer_bind     complement(1292..1314)
                     /label=Fragment.REV
     primer_bind     1297..1335
                     /label=Cherry delfwd 1
     misc_feature    1324
                     /label=5'
                     /note="5'"
     misc_feature    1336..1362
                     /label=AAAGAGGAA
     CDS             1378..1695
                     /codon_start=1
                     /label=FKBP
                     /note="FKBP"
                     /translation="VQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPF
                     KFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLK
                     LE"
     primer_bind     complement(1393..1416)
                     /label=FKBP-R
     primer_bind     1658..1680
                     /label=FKBP-F
     misc_feature    1702..1755
                     /label=GGSAx4GG
     primer_bind     complement(1765..1786)
                     /label=BFP-FKBP_vR
     misc_feature    1766..1782
                     /label=MCS
                     /note="3'"
     misc_feature    1771..1797
                     /label=MCS
                     /note="MCS"
     CDS             1798..3721
                     /codon_start=1
                     /label=PIP5K1C
                     /note="untitled_14"
                     /translation="MELEVPDEAESAEAGAVPSEAAWAAESGAAAGLAQKKAAPTEVLS
                     MTAQPGPGHGKKLGHRGVDASGETTYKKTTSSTLKGAIQLGIGYTVGHLSSKPERRVLM
                     QDFYVVESIFFPSEGSNLTPAHHFQDFRFKTYAPVAFRYFRELFGIRPDDYLYSLCNEP
                     LIELSNPGASGSLFYVTSDDEFIIKTVMHKEAEFLQKLLPGYYMNLNQNPRTLLPKFYG
                     LYCVQSGGKNIRVVVMNNILPRVVKMHLKFDLKGSTYKRRASKKEKEKSFPTYKDLDFM
                     QDMPEGLLLDADTFSALVKTLQDDCLVLESFKIMDYSLLLGVHNIDQHERERQAQGAQS
                     TSDEKRPVGQKALYSTAMESIQGGAARGEAIESDDTMGGIPAVNGRGERLLLHIGIIDI
                     LQSYRFIKKLEHTWKALVHDGDTVSVHRPSFYAERFFKFMSNTVFEENSSLKSSPSKKG
                     RGGALLAVKPLGPTAAFSASQIPSEREEAQYDLRGARSYPTLEDEGRPDLLPCTPPSFE
                     EATTASIATTLSSTSLSIPERSPSETSEQPRYRRRTQSSGQDGRPQEEPPAEEDLQQIT
                     VQVEPACSVEIVVPKEEDAGVEASPAGASAAVEVETASQASDEEGAPASQASDEEDAPA
                     TDIYF"
     CDS             2032..3129
                     /codon_start=1
                     /label=PIPK
                     /translation="LKGAIQLGIGYTVGHLSSKPERRVLMQDFYVVESIFFPSEGSNLT
                     PAHHFQDFRFKTYAPVAFRYFRELFGIRPDDYLYSLCNEPLIELSNPGASGSLFYVTSD
                     DEFIIKTVMHKEAEFLQKLLPGYYMNLNQNPRTLLPKFYGLYCVQSGGKNIRVVVMNNI
                     LPRVVKMHLKFDLKGSTYKRRASKKEKEKSFPTYKDLDFMQDMPEGLLLDADTFSALVK
                     TLQDDCLVLESFKIMDYSLLLGVHNIDQHERERQAQGAQSTSDEKRPVGQKALYSTAME
                     SIQGGAARGEAIESDDTMGGIPAVNGRGERLLLHIGIIDILQSYRFIKKLEHTWKALVH
                     DGDTVSVHRPSFYAERFFKFMSNTVF"
     primer_bind     complement(2032..2051)
                     /label=PPIKFyve-VR
     primer_bind     2032..2051
                     /label=PPIKFyve-IF
     misc_feature    2098..2100
                     /label=D101R
                     /note="mutation as compared to HsPIP5K1C, stops 
                     dimerization"
     misc_feature    2707..2709
                     /label=R304D
                     /note="mutation as compared to HsPIP5K1C, stops 
                     dimerization"
     primer_bind     complement(3105..3129)
                     /label=PPIKFyve-IR
     primer_bind     3112..3135
                     /label=PPIKFyve-VF
     misc_feature    3130..3135
                     /label=R445_K446_EE
                     /note="mutation as compared to HsPIP5K1C, stops membrane 
                     localization"
     primer_bind     complement(3145..3162)
                     /label=PIP5KgammaT1-455-R
     primer_bind     3703..3740
                     /label=BFP-FKBP_vF-CTerm
     misc_feature    3721
                     /label=3'
                     /note="3'"
     misc_feature    3721
                     /label=3'
                     /note="3'"
     misc_feature    3722..3746
                     /label=MCS
                     /note="MCS"
     primer_bind     3722..3741
                     /label=BFP-FKBP_vF
     primer_bind     3722..3740
                     /label=BFP-FKBP_vF-Kinase
     primer_bind     3722..3740
                     /label=BFPBB_PIKfyve_F
     misc_feature    3729..3760
                     /label=MCS
                     /note="3'"
     primer_bind     complement(3759..3778)
                     /label=SV40-pArev
     primer_bind     3812..3834
                     /label=A42
     misc_feature    3981..4436
                     /label=f1
                     /note="f1"
     promoter        4498..4526
                     /label=Bacterial
                     /note="Bacterial"
     promoter        4610..4839
                     /label=SV40early
                     /note="SV40early"
     misc_feature    4777..4912
                     /label=SV40
                     /note="SV40"
     misc_feature    4961..5755
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    5991..6009
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    6340..6983
                     /label=pUC
                     /note="pUC"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
      661 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
      721 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
      781 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
      841 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
      901 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      961 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     1021 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     1081 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     1141 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     1201 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     1261 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattccgga
     1321 ctcagatctc gaagcgcggc cgcgggagca ggaggagcag ctcgagcggc gctgggagtg
     1381 caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc
     1441 gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga
     1501 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     1561 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     1621 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     1681 cttctaaaac tggaaagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     1741 ggtagtgctg gtggtccgcg agctcaagct tcgaattctg cagtcgacct gcaggcgatg
     1801 gagctggagg taccggacga ggcagagagc gctgaggcag gggctgtgcc ctcggaggca
     1861 gcttgggctg ctgagagcgg ggctgcagca ggtttggctc agaagaaggc ggccccaaca
     1921 gaggttctgt ccatgacggc acagccgggc cctggccatg ggaagaagtt gggccatcga
     1981 ggtgtggacg catccggcga aaccacctac aagaagacca cctcctccac cctgaagggt
     2041 gccatccagc tgggcatcgg ctacaccgtg ggccacctga gctccaagcc cgaacgccgc
     2101 gtgctcatgc aggacttcta cgtggtggag agcatcttct tccccagcga aggcagcaac
     2161 ctcacccccg cccaccactt ccaggacttc cgcttcaaga cctatgcacc tgtcgccttc
     2221 cgctacttcc gggagctctt tgggatccgg ccagatgatt acttgtactc cctgtgcaat
     2281 gagccgctga tcgagctgtc caacccgggc gccagtggct ccctcttcta cgtcaccagc
     2341 gacgacgagt tcatcatcaa gaccgtcatg cacaaggagg ccgagttcct gcagaagctg
     2401 ctccctggct actacatgaa cctcaaccag aacccgcgga cgctgctgcc caagttctat
     2461 gggctgtact gcgtgcagtc ggggggcaag aacatccgcg tcgtggtcat gaacaacatc
     2521 ctgccccgcg tggtcaagat gcacctcaag ttcgacctca agggctccac ctacaagcgg
     2581 cgcgccagca agaaggagaa ggagaagagc ttccccacct acaaggacct ggacttcatg
     2641 caggacatgc ccgaggggct cctgctggac gccgacacct tcagcgccct ggtcaagacg
     2701 ctgcaggacg actgcctggt cctggaaagt ttcaagatca tggactacag cctgctgctg
     2761 ggcgtgcaca acatcgacca gcacgagcgc gagcggcagg cgcagggcgc ccagagcacc
     2821 tcagatgaga agcggcctgt gggccagaag gcgctctact ccacggccat ggagtccatc
     2881 cagggtggcg cagctcgcgg ggaggcaatc gaatcggatg acacgatggg cgggatcccc
     2941 gctgtgaacg gccgcgggga gcggctgctg ctgcacattg gcatcatcga catcctgcag
     3001 tcctacaggt tcatcaagaa actggagcac acctggaagg ccctcgtcca cgatggggac
     3061 acggtgtccg tccaccgccc cagcttctat gccgagcgct ttttcaagtt catgagcaac
     3121 acggtctttg aggagaactc ctccctgaag tcctcgccct ccaagaaggg gcgcggcgga
     3181 gccttgctag ctgtgaaacc actaggacct accgctgcat tctctgcaag tcagatccct
     3241 agtgaacgag aggaggctca gtacgacctt agaggggccc gcagttaccc cacgctggaa
     3301 gacgaaggcc gtcctgacct cctgccttgc acgccacctt ctttcgaaga agccactaca
     3361 gcctccattg ccacgactct gtcatccaca tccctctcca ttcctgaacg gtccccttcg
     3421 gaaacgtctg aacagccacg gtacaggcgg cgcacacagt catctggaca ggatggcagg
     3481 ccgcaggagg agccacccgc ggaagaggat ctgcagcaga ttacagtgca ggtggagcct
     3541 gcgtgcagcg tggagattgt ggtccccaaa gaggaggacg caggggtgga ggcttccccg
     3601 gccggtgcct ctgctgctgt tgaagtagaa actgccagcc aggcctcaga cgaggagggc
     3661 gcacctgcaa gccaggcatc ggacgaggag gacgcgcccg ccaccgacat ctacttttga
     3721 gcgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     3781 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3841 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3901 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3961 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4021 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4081 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4141 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4201 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4261 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4321 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4381 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     4441 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4501 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4561 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4621 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4681 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4741 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4801 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4861 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4921 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4981 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5041 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5101 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     5161 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5221 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5281 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5341 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5401 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5461 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     5521 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5581 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5641 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5701 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5761 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5821 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5881 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     5941 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     6001 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     6061 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     6121 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     6181 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     6241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6481 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7021 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
