LOCUS       sequence-207680-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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: mCherry-FKBP-SAC1
FEATURES             Location/Qualifiers
     source          1..6879
                     /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             1380..1700
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3666..3787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3703..3722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3757..3776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3794..4249)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3931..3952)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4143..4162
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4276..4380
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4382..4739
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4382..4573
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4405..4425)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4590..4725
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4774..5568
                     /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             4774..5568
                     /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(4828..4847)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5438..5457
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5756..5775)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5800..5847
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6176..6764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6665..6684
                     /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 atctcgaagc gcggccgcgg gagcaggagg agcagctcga gcggcgctgg
     1381 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     1441 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg
     1501 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     1561 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     1621 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     1681 tggagcttct aaaactggaa agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg
     1741 ctggtggtag tgctggtggt ccgcgagctc aagcttcgaa ttctgcaatg gcgacggcgg
     1801 cctacgagca gctgaagctg catatcacac ctgaaaaatt ttatgtggaa gcttgtgatg
     1861 atggagcaga tgacgtactt accattgacc gtgtgtccac agaggttacc cttgcagtca
     1921 agaaagatgt tcctccttca gctgtcacaa gaccaatatt tggtatactg ggcacaatcc
     1981 atctggtggc aggtaattat cttatagtca ttaccaaaaa gataaaagta ggtgaatttt
     2041 tcagtcatgt agtctggaaa gcaacagatt ttgatgtcct ttcttataag aagacaatgt
     2101 tgcacttaac tgatattcag ttacaagata ataaaacctt cctagcgatg ctaaaccatg
     2161 tcttgaatgt ggatggattt tacttttcaa caacatatga tttgacccat actttgcagc
     2221 ggctatccaa cactagtcct gaattccaag aaatgagtct cttggaaagg gcagatcagc
     2281 ggtttgtatg gaatggtcat cttctaagag aactttctgc acagccagag gttcatcggt
     2341 ttgcccttcc agtgttacat ggctttatta ccatgcattc atgttctatt aatggaaaat
     2401 actttgattg gattctcatc tcgaggagga gctgtttcag agctggtgtg cgctattatg
     2461 taagaggaat tgattcggaa ggccatgcag ctaactttgt agaaacagaa caaattgtgc
     2521 actacaatgg gagcaaagct tcgtttgtac agactcgagg atcaatacct gttttctggt
     2581 cccaaagacc aaacctcaag tacaaaccac tgccacagat cagcaaagta gcaaatcaca
     2641 tggacggttt ccaaaggcat tttgattccc aagtaattat ttatggaaaa caagttataa
     2701 tcaatctgat taaccagaag ggctcggaga agccacttga gcagacattt gcaacaatgg
     2761 tgtcttcctt gggaagtgga atgatgagat acattgcctt tgacttccat aaggaatgta
     2821 aaaatatgag atgggatcga ctaagtattt tattggatca ggtagcagaa atgcaagatg
     2881 aattaagtta ttttctagtg gactctgctg gccaggtggt ggcaaaccag gaaggcgtgt
     2941 tccgaagcaa ttgcatggat tgtctagata gaaccaatgt gatccagagt ttgttagctc
     3001 gtcgttcact tcaggcccaa cttcagagac taggagtttt gcatgtggga caaaagcttg
     3061 aagaacaaga tgaatttgag aagattttca aaaatgcctg ggctgacaac gcaaatgctt
     3121 gtgccaagca atatgcggga actggtgcct tgaagactga ctttaccaga actggaaaga
     3181 gaactcattt gggacttata atggatggct ggaactcaat gatacgatat tataagaaca
     3241 acttttccga tggatttaga caagattcca tagacttatt tcttggaaac tattcagtgg
     3301 atgaattaga atctcatagt cctttaagtg ttccaaggga ctggaaattc ctggctttgc
     3361 ctattatcat ggttgttgcc ttttcaatgt gcattatctg tttgcttatg gctggtgaca
     3421 cttggacaga aacactggcc tatgtgctct tctggggagt tgcaagcatt ggaacatttt
     3481 ttatcattct ttacaatggc aaagattttg tcgatgctcc cagactggtc cagaaagaaa
     3541 agatagactg aagataactg atcataatca gccataccac atttgtagag gttttacttg
     3601 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3661 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3721 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3781 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3841 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3901 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3961 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4021 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4081 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4141 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4201 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4261 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4321 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4381 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4441 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4501 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4561 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4621 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4681 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4741 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5281 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5581 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5641 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5701 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5761 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5821 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5881 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5941 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6001 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6841 cccctgattc tgtggataac cgtattaccg ccatgcatt
//
