LOCUS       sequence-203963-        5328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI4P biosensor .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5328)
  AUTHORS   Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR
  TITLE     SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic 
            reticulum to maintain a steep chemical gradient with donor 
            membranes.
  JOURNAL   Elife. 2018 Feb 20;7. pii: 35588. doi: 10.7554/eLife.35588.
  PUBMED    29461204
REFERENCE   2  (bases 1 to 5328)
  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: "Elife."; 
            date: "2018-02-20"; volume: "7. pii: 35588. doi"; pages: " 
            10.7554/eLife.35588"
COMMENT     Sequence Label: NES-iRFP-P4Mx1
FEATURES             Location/Qualifiers
     source          1..5328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             658..1605
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     CDS             1678..1983
                     /codon_start=1
                     /product="high-affinity phosphatidylinositol 4-phosphate 
                     binding module of SidM (DrrA) from Legionella pneumophila 
                     (Hammond et al., 2014)"
                     /label=P4M
                     /translation="TASTENFKNVKEKYQQMRGDALKTEILADFKDKLAEATDEQSLKQ
                     IVAELKSKDEYRILAKGQGLTTQLLGLKTSSVSSFEKMVEETRESIKSQERQTIKIK"
     polyA_signal    2116..2237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2153..2172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2207..2226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2244..2699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2381..2402)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2593..2612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2726..2830
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2832..3189
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2832..3023
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2855..2875)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3040..3175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3224..4018
                     /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             3224..4018
                     /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(3278..3297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3888..3907
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4206..4225)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4250..4297
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4626..5214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5115..5134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gctagccacc
      601 atggctctgc agaaaaagtt ggaagagctt gagctggatg aggcaccggt cgccaccatg
      661 gcggaaggat ccgtcgccag gcagcctgac ctcttgacct gcgacgatga gccgatccat
      721 atccccggtg ccatccaacc gcatggactg ctgctcgccc tcgccgccga catgacgatc
      781 gttgccggca gcgacaacct tcccgaactc accggactgg cgatcggcgc cctgatcggc
      841 cgctctgcgg ccgatgtctt cgactcggag acgcacaacc gtctgacgat cgccttggcc
      901 gagcccgggg cggccgtcgg agcaccgatc actgtcggct tcacgatgcg aaaggacgca
      961 ggcttcatcg gctcctggca tcgccatgat cagctcatct tcctcgagct cgagcctccc
     1021 cagcgggacg tcgccgagcc gcaggcgttc ttccgccgca ccaacagcgc catccgccgc
     1081 ctgcaggccg ccgaaacctt ggaaagcgcc tgcgccgccg cggcgcaaga ggtgcggaag
     1141 attaccggct tcgatcgggt gatgatctat cgcttcgcct ccgacttcag cggcgaagtg
     1201 atcgcagagg atcggtgcgc cgaggtcgag tcaaaactag gcctgcacta tcctgcctca
     1261 accgtgccgg cgcaggcccg tcggctctat accatcaacc cggtacggat cattcccgat
     1321 atcaattatc ggccggtgcc ggtcacccca gacctcaatc cggtcaccgg gcggccgatt
     1381 gatcttagct tcgccatcct gcgcagcgtc tcgcccgtcc atctggaatt catgcgcaac
     1441 ataggcatgc acggcacgat gtcgatctcg attttgcgcg gcgagcgact gtggggattg
     1501 atcgtttgcc atcaccgaac gccgtactac gtcgatctcg atggccgcca agcctgcgag
     1561 ctagtcgccc aggttctggc ctggcagatc ggcgtgatgg aagagtccgg actcagatct
     1621 cgagctcaag cttcgaattc tgcagtcgac ggtggtagtg cttccggact cagatccacg
     1681 gcaagcacgg aaaactttaa aaatgttaaa gaaaaatatc agcaaatgcg aggtgatgct
     1741 ttaaaaacag aaatcctggc tgatttcaag gataaactgg ctgaagctac cgatgaacag
     1801 agccttaagc aaattgttgc tgaattgaaa agtaaagatg aatatagaat tttggctaag
     1861 gggcaagggt taacaaccca gcttttaggg ttaaagacaa gttcagtgtc ttcatttgag
     1921 aaaatggttg aggaaacaag agaaagtatt aagtctcaag aaagacaaac cattaagata
     1981 aaataaggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     2041 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2101 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2221 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     2281 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2341 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2401 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2461 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2521 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2581 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2641 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2701 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2761 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2821 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2881 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2941 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3001 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3061 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3121 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3181 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3241 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3301 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3361 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3421 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3481 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3541 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3601 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3661 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3721 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3781 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3841 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3901 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3961 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4021 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4081 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4141 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4201 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4261 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4321 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4381 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4441 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4501 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4741 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5281 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
