LOCUS       sequence-207691-        6792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable inactive SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6792)
  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 6792)
  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-SAC1C389S-FKBP
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             611..1318
                     /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(756..774)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1024..1043
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3134..3451
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="VQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPF
                     KFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLK
                     LE"
     polyA_signal    3578..3699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3615..3634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3669..3688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3706..4161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3843..3864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4055..4074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4188..4292
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4294..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4294..4485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4686..5480
                     /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             4686..5480
                     /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(4740..4759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5350..5369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5668..5687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5712..5759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6088..6676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6577..6596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      601 ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
      661 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
      721 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
      781 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
      841 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
      901 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
      961 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1021 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     1081 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     1141 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     1201 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1261 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc
     1321 cggactcaga tctatggcga cggcggccta cgagcagctg aagctgcata tcacacctga
     1381 aaaattttat gtggaagctt gtgatgatgg agcagatgac gtacttacca ttgaccgtgt
     1441 gtccacagag gttacccttg cagtcaagaa agatgttcct ccttcagctg tcacaagacc
     1501 aatatttggt atactgggca caatccatct ggtggcaggt aattatctta tagtcattac
     1561 caaaaagata aaagtaggtg aatttttcag tcatgtagtc tggaaagcaa cagattttga
     1621 tgtcctttct tataagaaga caatgttgca cttaactgat attcagttac aagataataa
     1681 aaccttccta gcgatgctaa accatgtctt gaatgtggat ggattttact tttcaacaac
     1741 atatgatttg acccatactt tgcagcggct atccaacact agtcctgaat tccaagaaat
     1801 gagtctcttg gaaagggcag atcagcggtt tgtatggaat ggtcatcttc taagagaact
     1861 ttctgcacag ccagaggttc atcggtttgc ccttccagtg ttacatggct ttattaccat
     1921 gcattcatgt tctattaatg gaaaatactt tgattggatt ctcatctcga ggaggagctg
     1981 tttcagagct ggtgtgcgct attatgtaag aggaattgat tcggaaggcc atgcagctaa
     2041 ctttgtagaa acagaacaaa ttgtgcacta caatgggagc aaagcttcgt ttgtacagac
     2101 tcgaggatca atacctgttt tctggtccca aagaccaaac ctcaagtaca aaccactgcc
     2161 acagatcagc aaagtagcaa atcacatgga cggtttccaa aggcattttg attcccaagt
     2221 aattatttat ggaaaacaag ttataatcaa tctgattaac cagaagggct cggagaagcc
     2281 acttgagcag acatttgcaa caatggtgtc ttccttggga agtggaatga tgagatacat
     2341 tgcctttgac ttccataagg aatgtaaaaa tatgagatgg gatcgactaa gtattttatt
     2401 ggatcaggta gcagaaatgc aagatgaatt aagttatttt ctagtggact ctgctggcca
     2461 ggtggtggca aaccaggaag gcgtgttccg aagcaatagc atggattgtc tagatagaac
     2521 caatgtgatc cagagtttgt tagctcgtcg ttcacttcag gcccaacttc agagactagg
     2581 agttttgcat gtgggacaaa agcttgaaga acaagatgaa tttgagaaga ttttcaaaaa
     2641 tgcctgggct gacaacgcaa atgcttgtgc caagcaatat gcgggaactg gtgccttgaa
     2701 gactgacttt accagaactg gaaagagaac tcatttggga cttataatgg atggctggaa
     2761 ctcaatgata cgatattata agaacaactt ttccgatgga tttagacaag attccataga
     2821 cttatttctt ggaaactatt cagtggatga attagaatct catagtcctt taagtgttcc
     2881 aagggactgg aaattcctgg ctttgcctat tatcatggtt gttgcctttt caatgtgcat
     2941 tatctgtttg cttatggctg gtgacacttg gacagaaaca ctggcctatg tgctcttctg
     3001 gggagttgca agcattggaa cattttttat cattctttac aatggcaaag attttgtcga
     3061 tgctcccaga ctggtccaga aagaaaagat agacggaatt tgtatttgtg gaaagcggct
     3121 tggcttgggt accgtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa
     3181 gcgcggccag acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga
     3241 ttcctcccgg gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg
     3301 aggctgggaa gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc
     3361 tccagattat gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct
     3421 cgtcttcgat gtggagcttc taaaactgga atccaccgga tctagataac tgatcataat
     3481 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3541 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3721 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3781 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3841 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3901 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3961 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4021 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4081 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4141 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4261 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4681 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4741 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4801 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4861 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4921 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4981 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5041 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5101 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5161 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5221 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5281 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5341 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5401 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5461 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5521 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5581 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5641 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5701 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5761 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5821 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5881 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5941 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6001 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6061 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6241 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6721 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6781 cgccatgcat ta
//
