LOCUS       sequence-434121-        6406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses ORP5 with the OSBP-related ligand-binding domain and the 
            transmembrane domain deleted. The mCherry fluorophore and FKBP are 
            attached to ORP5. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  AUTHORS   Doyle CP, Rectenwald A, Timple L, Hammond GRV
  TITLE     Orthogonal Targeting of SAC1 to Mitochondria Implicates ORP2 as a 
            Major Player in PM PI4P Turnover.
  JOURNAL   Contact (Thousand Oaks). 2024 Feb 7;7:25152564241229272. doi: 
            10.1177/25152564241229272. eCollection 2024 Jan-Dec.
  PUBMED    38327560
REFERENCE   2  (bases 1 to 6406)
  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: "Contact 
            (Thousand Oaks)."; date: "2024-02-7"; volume: "7:25152564241229272. 
            doi"; pages: " 10.1177/25152564241229272. eCollection 2024 Jan-Dec"
COMMENT     Sequence Label: mCherry-C1-ORP5deltaORDdeltaTMD-FKBP
FEATURES             Location/Qualifiers
     source          1..6406
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(2737..2739,2740..3060)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    3194..3315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3231..3250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3285..3304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3322..3777)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3459..3480)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3671..3690
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3804..3908
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3910..4267
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3910..4101
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      4118..4253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4302..5096
                     /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             4302..5096
                     /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(4356..4375)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4966..4985
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5284..5303)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5328..5375
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5704..6292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6193..6212
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc gaaggaggag gccttcctcc ggcgccgctt ctccctgtgt
     1381 ccaccttcct ccacccctca gaaagtcgac ccccggaagc tcacccggaa cttgctcctc
     1441 agcggagaca atgagctcta cccactcagc ccagggaagg acatggagcc caacggcccg
     1501 tcgctgccca gggatgaagg gcccccgacc ccaagctctg ccacgaaggt gccaccggca
     1561 gagtacaggc tgtgcaacgg gtcagacaag gaatgtgtgt cccccaccgc cagggtcacc
     1621 aagaaggaga ctctcaaggc gcagaaggag aactaccggc aggagaagaa gcgcgccaca
     1681 cggcagctgc tcagcgctct gacagacccc agcgtggtca tcatggctga cagcctgaag
     1741 atccgcggca ccctgaagag ctggaccaag ctgtggtgcg tgctgaagcc gggggtgctg
     1801 ctcatctaca agacgcccaa ggtgggccag tgggtgggca cggtgctgct gcactgctgc
     1861 gagctcatcg agcggccctc caagaaggac ggcttctgct tcaagctctt ccacccgctg
     1921 gatcagtccg tctgggccgt gaagggcccc aaaggtgaga gcgtgggctc catcacacag
     1981 cccctgccca gcagctacct gatcttcagg gccgcctccg agtcagatgg tcgctgctgg
     2041 ctggacgccc tggagctggc cctgcgctgc tctagcctac tgagactggg cacctgcaag
     2101 ccgggccgag acggggagcc agggacctcg ccagacgcat caccctcatc gctctgtggg
     2161 ctgccagcct cagccaccgt ccacccagac caagacctgt tcccactgaa cgggtcttcc
     2221 ctggagaacg atgcattctc agacaagtcg gagagagaga accctgagga gtcagatacc
     2281 gagacccagg accatagccg gaagacggag agtggcagcg accagtcaga gacccctggg
     2341 gccccggtgc ggagaggagg aagcggagga caggaggccg tggcccgcca gaccaccttc
     2401 ctgggcagcc cagggcccag gcacgagagg tctggcccag accagcggct tcgcaaggcc
     2461 agcgaccagc cctccggcca cagccaggcc acggagagca gcggatccac gcctgagtcc
     2521 tgcccagagc tctcagacga ggagcaggat ggtgactttg tccctggcgg tgagagccca
     2581 tgccctcggt gcaggaagga ggcgcggcgg ctgcaggccc tgcacgaggc catcctctcc
     2641 atccgagagg cccagcagga gctgcacagg cacctctcgg ccatgctgag ctccacggca
     2701 cgggcagcac aggcaccgac cccaggcctc ctgcagatgg gagtgcaggt ggaaaccatc
     2761 tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt gcactacacc
     2821 gggatgcttg aagatggaaa gaaatttgat tcctcccggg acagaaacaa gccctttaag
     2881 tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt
     2941 gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac
     3001 ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct aaaactggaa
     3061 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3121 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3181 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3241 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3301 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3361 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3421 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3481 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3541 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3601 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3661 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3721 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3781 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3841 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3901 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3961 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4021 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4081 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4141 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4201 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4261 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4321 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4381 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4441 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4501 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4561 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4621 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4681 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4741 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4801 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4861 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4921 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4981 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5041 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5101 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5161 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5221 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5281 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5341 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5401 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5461 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5521 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5581 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5641 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5821 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5941 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6001 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6061 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6121 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6181 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6241 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6301 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6361 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
