LOCUS       sequence-207682-        6879 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 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-SAC1C389S
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        46..349
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        350..553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     504..524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      592..601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             598..1305
                     /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(743..761)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1011..1030
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1366..1686
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3652..3773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3689..3708)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3743..3762
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3780..4235)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3917..3938)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4129..4148
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4262..4366
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4368..4725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4368..4559
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4391..4411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4576..4711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4760..5554
                     /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             4760..5554
                     /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(4814..4833)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5424..5443
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5742..5761)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5786..5833
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6162..6750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6651..6670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
       61 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      121 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      181 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      241 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      301 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      361 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      421 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      481 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      541 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
      601 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
      661 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      721 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
      781 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
      841 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      901 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      961 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1021 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     1081 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     1141 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     1201 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     1261 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtccgg actcagatct
     1321 cgaagcgcgg ccgcgggagc aggaggagca gctcgagcgg cgctgggagt gcaggtggaa
     1381 accatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac
     1441 tacaccggga tgcttgaaga tggaaagaaa tttgattcct cccgggacag aaacaagccc
     1501 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
     1561 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
     1621 gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa
     1681 ctggaaagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     1741 ggtggtccgc gagctcaagc ttcgaattct gcaatggcga cggcggccta cgagcagctg
     1801 aagctgcata tcacacctga aaaattttat gtggaagctt gtgatgatgg agcagatgac
     1861 gtacttacca ttgaccgtgt gtccacagag gttacccttg cagtcaagaa agatgttcct
     1921 ccttcagctg tcacaagacc aatatttggt atactgggca caatccatct ggtggcaggt
     1981 aattatctta tagtcattac caaaaagata aaagtaggtg aatttttcag tcatgtagtc
     2041 tggaaagcaa cagattttga tgtcctttct tataagaaga caatgttgca cttaactgat
     2101 attcagttac aagataataa aaccttccta gcgatgctaa accatgtctt gaatgtggat
     2161 ggattttact tttcaacaac atatgatttg acccatactt tgcagcggct atccaacact
     2221 agtcctgaat tccaagaaat gagtctcttg gaaagggcag atcagcggtt tgtatggaat
     2281 ggtcatcttc taagagaact ttctgcacag ccagaggttc atcggtttgc ccttccagtg
     2341 ttacatggct ttattaccat gcattcatgt tctattaatg gaaaatactt tgattggatt
     2401 ctcatctcga ggaggagctg tttcagagct ggtgtgcgct attatgtaag aggaattgat
     2461 tcggaaggcc atgcagctaa ctttgtagaa acagaacaaa ttgtgcacta caatgggagc
     2521 aaagcttcgt ttgtacagac tcgaggatca atacctgttt tctggtccca aagaccaaac
     2581 ctcaagtaca aaccactgcc acagatcagc aaagtagcaa atcacatgga cggtttccaa
     2641 aggcattttg attcccaagt aattatttat ggaaaacaag ttataatcaa tctgattaac
     2701 cagaagggct cggagaagcc acttgagcag acatttgcaa caatggtgtc ttccttggga
     2761 agtggaatga tgagatacat tgcctttgac ttccataagg aatgtaaaaa tatgagatgg
     2821 gatcgactaa gtattttatt ggatcaggta gcagaaatgc aagatgaatt aagttatttt
     2881 ctagtggact ctgctggcca ggtggtggca aaccaggaag gcgtgttccg aagcaatagc
     2941 atggattgtc tagatagaac caatgtgatc cagagtttgt tagctcgtcg ttcacttcag
     3001 gcccaacttc agagactagg agttttgcat gtgggacaaa agcttgaaga acaagatgaa
     3061 tttgagaaga ttttcaaaaa tgcctgggct gacaacgcaa atgcttgtgc caagcaatat
     3121 gcgggaactg gtgccttgaa gactgacttt accagaactg gaaagagaac tcatttggga
     3181 cttataatgg atggctggaa ctcaatgata cgatattata agaacaactt ttccgatgga
     3241 tttagacaag attccataga cttatttctt ggaaactatt cagtggatga attagaatct
     3301 catagtcctt taagtgttcc aagggactgg aaattcctgg ctttgcctat tatcatggtt
     3361 gttgcctttt caatgtgcat tatctgtttg cttatggctg gtgacacttg gacagaaaca
     3421 ctggcctatg tgctcttctg gggagttgca agcattggaa cattttttat cattctttac
     3481 aatggcaaag attttgtcga tgctcccaga ctggtccaga aagaaaagat agactgaaga
     3541 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3601 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3661 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3721 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     3781 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3841 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3901 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3961 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     4021 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4081 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4141 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4201 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4261 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4321 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     4381 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4501 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4561 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4621 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4681 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4741 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4801 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4861 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4921 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4981 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5041 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5101 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5161 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5221 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5281 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5341 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5401 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5461 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5521 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5581 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5641 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5701 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5761 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5821 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5881 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5941 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     6001 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6301 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6781 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6841 gataaccgta ttaccgccat gcattagtta ttaatagta
//
