LOCUS       sequence-434109-        7582 bp    DNA     circular SYN 15-APR-2025
DEFINITION  siRNA resistant ORP5 attcahed with anFKBP and a mCherry fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  AUTHORS   Doyle CP, Timple L, Hammond GRV
  TITLE     OSBP is a Major Determinant of Golgi Phosphatidylinositol 
            4-Phosphate Homeostasis.
  JOURNAL   Contact (Thousand Oaks). 2024 Feb 22;7:25152564241232196. doi: 
            10.1177/25152564241232196. eCollection 2024 Jan-Dec.
  PUBMED    38405037
REFERENCE   2  (bases 1 to 7582)
  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: "Contact 
            (Thousand Oaks)."; date: "2024-02-22"; volume: "7:25152564241232196.
            doi"; pages: " 10.1177/25152564241232196. eCollection 2024 Jan-Dec"
COMMENT     Sequence Label: mCherry-C1-FKBPflex-OSBP-siRNAr
FEATURES             Location/Qualifiers
     source          1..7582
                     /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             1381..1701
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    4370..4491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4407..4426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4461..4480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4498..4953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4635..4656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4847..4866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4980..5084
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5086..5443
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5086..5277
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5109..5129)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5294..5429
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5478..6272
                     /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             5478..6272
                     /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(5532..5551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6142..6161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6460..6479)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6504..6551
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6880..7468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7369..7388
                     /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 gatctcgaag cgcggccgcg ggagcaggag gagcagctcg agcggcgctg
     1381 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag
     1441 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
     1501 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
     1561 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
     1621 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
     1681 gtggagcttc taaaactgga aagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     1741 gctggtggta gtgctggtgg tccgcgagct caagcttcga attctatggc ggcaactgaa
     1801 ttgcggggtg ttgtgggacc cggtcccgcg gctattgccg cactcggagg tggaggggcc
     1861 ggcccgcccg ttgtaggagg aggtggcgga cgaggtgacg cagggcctgg aagcggtgct
     1921 gctagtggca ccgtagtagc agcagcagcg ggtggccccg gccctggagc tggaggtgtt
     1981 gctgcagcag gaccagcacc ggctcctcca acggggggga gcggaggatc tggagcgggg
     2041 ggcagtggat ctgccaggga agggtggttg ttcaagtgga ccaactatat aaaaggttac
     2101 caacggagat ggtttgtcct gagtaacggc ctcctttcct actaccgaag taaggcagaa
     2161 atgcgccata cttgtcgcgg aacaattaat ttggccactg cgaatataac ggtagaggat
     2221 agctgtaact ttattatcag caacggggga gcacagacgt accacctgaa agcatcatca
     2281 gaggttgagc ggcaaaggtg ggtcaccgca ctggagctcg cgaaggccaa ggcggtgaaa
     2341 atgctggcag agagtgacga gtctggggat gaggagagcg tctctcagac tgataagacc
     2401 gagttgcaga atacattgcg aacccttagc tcaaaagtcg aggatctgag cacctgtaac
     2461 gacttgatag ctaaacatgg caccgccctt cagaggtcac tgtctgagct tgaaagtctc
     2521 aagcttccag cagagtccaa tgagaagatt aagcaggtca atgaaagggc gacgctcttc
     2581 cggataacga gtaatgctat gattaacgcg tgcagggact tccttatgct ggcccagaca
     2641 cacagcaaaa aatggcaaaa atcactgcag tacgagcggg accaacgcat acgacttgaa
     2701 gaaacattgg agcagctggc caagcaacat aaccatttgg aaagggcttt taggggagca
     2761 acagtcctcc ccgcaaatac tccgggcaac gtcggctccg gcaaggatca atgctgctcc
     2821 ggtaaaggtg atatgtctga cgaagacgat gaaaatgaat tcttcgatgc ccctgaaatt
     2881 attacaatgc cagagaacct cggacacaaa cgaacaggca gtaatatatc aggggccagc
     2941 tctgatatat cattggatga acagtataaa catcagttgg aagaaacaaa aaaggagaaa
     3001 agaacccgga ttccatacaa accaaactat tcactcaact tgtggtctat tatgaagaat
     3061 tgcatcggga aggagttgtc aaagatccca atgccagtca acttcaacga acctctgtcc
     3121 atgcttcaga gactgaccga ggacttggag tatcacgagt tgttggacag ggccgctaag
     3181 tgtgagaatt cattggagca actttgttat gtggctgctt ttactgtcag ctcttacagc
     3241 actactgtat ttcgcacgtc taaaccattc aacccactgc ttggggaaac attcgaactg
     3301 gacagattgg aagagaatgg ctaccgaagc ctttgtgaac aggttagtca tcatccgcca
     3361 gcagccgccc accatgcaga gtccaagaac ggctggaccc tgcgacagga gataaaaatc
     3421 acgtccaaat ttagaggaaa atatctctca ataatgcctc ttggaaccat acactgcata
     3481 ttccatgcca ctggtcacca ttatacatgg aaaaaagtga cgacgacagt gcataacatc
     3541 atagtcggta aactgtggat cgaccaatca ggagaaatag atatagtgaa ccacaaaacg
     3601 ggggataagt gcaatctcaa gtttgtaccg tactcttatt tttcacgaga tgtcgcaagg
     3661 aaagttacgg gtgaagtaac ggacccgtca ggcaaagtac attttgccct gctcgggact
     3721 tgggatgaga aaatggagtg ctttaaggta cagccggtta ttggtgagaa tggaggggat
     3781 gcaagacaaa gaggtcacga agccgaggaa tcacgggtaa tgctgtggaa gagaaaccca
     3841 ttgccgaaga acgcagagaa catgtattat ttctcagaat tggcccttac cctgaacgcc
     3901 tgggagtcag gaaccgcgcc cacggactca cgacttagac cggaccaaag attgatggag
     3961 aacggacggt gggatgaggc aaacgcagag aagcagcgac tggaggagaa gcaacgattg
     4021 tccagaaaaa agcgcgaggc tgaagctatg aaagctacag aagatggtac tccctatgac
     4081 ccttacaagg ccctctggtt cgagcggaag aaagacccgg taaccaagga gctcacccat
     4141 atttatcgcg gggagtattg ggagtgcaag gaaaagcagg attggtccag ctgtccagat
     4201 atcttctgag aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata
     4261 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4321 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4381 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4441 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     4501 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4561 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4621 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4681 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4741 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4801 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4861 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4921 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     5101 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5161 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5221 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5281 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5341 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5401 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     5461 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5521 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5581 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5641 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5701 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5761 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5821 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5881 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5941 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6001 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6061 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6121 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6181 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6241 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     6301 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6361 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6421 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     6481 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     6541 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6601 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6661 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6721 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6781 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6841 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6901 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6961 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7021 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7081 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7141 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7201 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7261 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7321 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7381 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7441 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7501 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7561 taaccgtatt accgccatgc at
//
