LOCUS       sequence-434120-        7576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses ORP5 with 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 7576)
  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 7576)
  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-ORP5deltaTMD-FKBP
FEATURES             Location/Qualifiers
     source          1..7576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             612..1319
                     /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(757..775)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1025..1044
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(3906..3908,3909..4229)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    4363..4484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4400..4419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4454..4473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4491..4946)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4628..4649)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4840..4859
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4973..5077
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5079..5436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5079..5270
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      5287..5422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5471..6265
                     /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             5471..6265
                     /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(5525..5544)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6135..6154
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6453..6472)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6497..6544
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6873..7461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7362..7381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
      661 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      721 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      781 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      841 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      901 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      961 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1021 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1081 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1141 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1201 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1261 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     1321 ccggactcag atctcgagcg aaggaggagg ccttcctccg gcgccgcttc tccctgtgtc
     1381 caccttcctc cacccctcag aaagtcgacc cccggaagct cacccggaac ttgctcctca
     1441 gcggagacaa tgagctctac ccactcagcc cagggaagga catggagccc aacggcccgt
     1501 cgctgcccag ggatgaaggg cccccgaccc caagctctgc cacgaaggtg ccaccggcag
     1561 agtacaggct gtgcaacggg tcagacaagg aatgtgtgtc ccccaccgcc agggtcacca
     1621 agaaggagac tctcaaggcg cagaaggaga actaccggca ggagaagaag cgcgccacac
     1681 ggcagctgct cagcgctctg acagacccca gcgtggtcat catggctgac agcctgaaga
     1741 tccgcggcac cctgaagagc tggaccaagc tgtggtgcgt gctgaagccg ggggtgctgc
     1801 tcatctacaa gacgcccaag gtgggccagt gggtgggcac ggtgctgctg cactgctgcg
     1861 agctcatcga gcggccctcc aagaaggacg gcttctgctt caagctcttc cacccgctgg
     1921 atcagtccgt ctgggccgtg aagggcccca aaggtgagag cgtgggctcc atcacacagc
     1981 ccctgcccag cagctacctg atcttcaggg ccgcctccga gtcagatggt cgctgctggc
     2041 tggacgccct ggagctggcc ctgcgctgct ctagcctact gagactgggc acctgcaagc
     2101 cgggccgaga cggggagcca gggacctcgc cagacgcatc accctcatcg ctctgtgggc
     2161 tgccagcctc agccaccgtc cacccagacc aagacctgtt cccactgaac gggtcttccc
     2221 tggagaacga tgcattctca gacaagtcgg agagagagaa ccctgaggag tcagataccg
     2281 agacccagga ccatagccgg aagacggaga gtggcagcga ccagtcagag acccctgggg
     2341 ccccggtgcg gagagggacc acctatgtgg agcaggtcca ggaggagctg ggggagctgg
     2401 gcgaggcgtc ccaggtggag acagtgtcag aggagaacaa gagtctgatg tggaccctgc
     2461 tgaagcagct acggccaggc atggacctgt cccgcgtggt gctacccacg ttcgtactgg
     2521 agccgcgctc cttcctgaac aagctctccg actactacta ccacgcagac ctgctctcca
     2581 gggctgcggt ggaggaggat gcctacagcc gcatgaagct ggtgctgcgg tggtacctgt
     2641 ctggcttcta caagaagccc aagggaatca agaagccgta caaccccatc ctgggggaga
     2701 ccttccgctg ctgctggttc cacccgcaga ctgacagccg cacattctac atagcagagc
     2761 aggtgtccca ccacccgccc gtgtctgcct tccacgtcag caaccggaag gacggcttct
     2821 gcatcagtgg cagcatcaca gccaagtcca ggttttatgg gaactcgctg tcggcgctgc
     2881 tggacggcaa agccacgctc accttcctga accgagccga ggattacacc cttaccatgc
     2941 cctacgccca ctgcaaagga atcctgtatg gcacgatgac cctggagctg ggtgggaagg
     3001 tcaccatcga gtgtgcgaag aacaacttcc aggcccagct ggaattcaaa ctcaagccct
     3061 tcttcggggg tagcaccagc atcaaccaga tctcgggaaa gatcacgtcg ggagaggaag
     3121 tcctggcgag cctcagtggc cactgggaca gggacgtgtt tatcaaggag gaagggagcg
     3181 gaagcagtgc gcttttctgg accccgagcg gggaggtccg cagacagagg ctgaggcagc
     3241 acacggtgcc gctggaggag cagacggagc tggagtccga gaggctctgg cagcacgtca
     3301 ccagggccat cagcaagggc gaccagcaca gggccacaca ggagaagttt gcactggagg
     3361 aggcacagcg gcagcgggcc cgtgagcggc aggagagcct catgccctgg aagccgcagc
     3421 tgttccacct ggaccccatc acccaggagt ggcactaccg atacgaggac cacagcccct
     3481 gggaccccct gaaggacatc gcccagtttg agcaagacgg gatcctgcgg accttgcagc
     3541 aggaggccgt ggcccgccag accaccttcc tgggcagccc agggcccagg cacgagaggt
     3601 ctggcccaga ccagcggctt cgcaaggcca gcgaccagcc ctccggccac agccaggcca
     3661 cggagagcag cggatccacg cctgagtcct gcccagagct ctcagacgag gagcaggatg
     3721 gtgactttgt ccctggcggt gagagcccat gccctcggtg caggaaggag gcgcggcggc
     3781 tgcaggccct gcacgaggcc atcctctcca tccgagaggc ccagcaggag ctgcacaggc
     3841 acctctcggc catgctgagc tccacggcac gggcagcaca ggcaccgacc ccaggcctcc
     3901 tgcagatggg agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc
     3961 gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaatttgatt
     4021 cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag
     4081 gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc
     4141 cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg
     4201 tcttcgatgt ggagcttcta aaactggaac ccgggatcca ccggatctag ataactgatc
     4261 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4321 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4381 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4441 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4501 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4561 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4621 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4681 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4741 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4801 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4861 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4921 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4981 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5041 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5101 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5161 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5281 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5341 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5401 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5461 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5521 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5581 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5641 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5701 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5761 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5821 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5881 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5941 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     6001 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6061 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6121 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6181 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6241 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6301 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6361 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6421 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6481 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6541 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6601 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6661 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7081 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7201 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7261 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7321 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7381 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7441 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7501 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7561 attaccgcca tgcatt
//
