LOCUS       sequence-207671-        6586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable cytosolic inactive SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  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 6586)
  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: SAC1C389SdeltaTMD-FKBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        370..573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2202..2204,2205..2525)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      2541..2550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2547..3257
                     /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(2692..2710)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2960..2979
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3379..3500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3416..3435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3470..3489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3507..3962)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3644..3665)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3856..3875
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3989..4093
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4095..4452
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4095..4286
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4118..4138)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4303..4438
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4487..5281
                     /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             4487..5281
                     /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(4541..4560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5151..5170
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5469..5488)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5513..5560
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5889..6477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6378..6397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      601 cgctaccgga ctcagatctc gagctcatgg cgacggcggc ctacgagcag ctgaagctgc
      661 atatcacacc tgaaaaattt tatgtggaag cttgtgatga tggagcagat gacgtactta
      721 ccattgaccg tgtgtccaca gaggttaccc ttgcagtcaa gaaagatgtt cctccttcag
      781 ctgtcacaag accaatattt ggtatactgg gcacaatcca tctggtggca ggtaattatc
      841 ttatagtcat taccaaaaag ataaaagtag gtgaattttt cagtcatgta gtctggaaag
      901 caacagattt tgatgtcctt tcttataaga agacaatgtt gcacttaact gatattcagt
      961 tacaagataa taaaaccttc ctagcgatgc taaaccatgt cttgaatgtg gatggatttt
     1021 acttttcaac aacatatgat ttgacccata ctttgcagcg gctatccaac actagtcctg
     1081 aattccaaga aatgagtctc ttggaaaggg cagatcagcg gtttgtatgg aatggtcatc
     1141 ttctaagaga actttctgca cagccagagg ttcatcggtt tgcccttcca gtgttacatg
     1201 gctttattac catgcattca tgttctatta atggaaaata ctttgattgg attctcatct
     1261 cgaggaggag ctgtttcaga gctggtgtgc gctattatgt aagaggaatt gattcggaag
     1321 gccatgcagc taactttgta gaaacagaac aaattgtgca ctacaatggg agcaaagctt
     1381 cgtttgtaca gactcgagga tcaatacctg ttttctggtc ccaaagacca aacctcaagt
     1441 acaaaccact gccacagatc agcaaagtag caaatcacat ggacggtttc caaaggcatt
     1501 ttgattccca agtaattatt tatggaaaac aagttataat caatctgatt aaccagaagg
     1561 gctcggagaa gccacttgag cagacatttg caacaatggt gtcttccttg ggaagtggaa
     1621 tgatgagata cattgccttt gacttccata aggaatgtaa aaatatgaga tgggatcgac
     1681 taagtatttt attggatcag gtagcagaaa tgcaagatga attaagttat tttctagtgg
     1741 actctgctgg ccaggtggtg gcaaaccagg aaggcgtgtt ccgaagcaat agcatggatt
     1801 gtctagatag aaccaatgtg atccagagtt tgttagctcg tcgttcactt caggcccaac
     1861 ttcagagact aggagttttg catgtgggac aaaagcttga agaacaagat gaatttgaga
     1921 agattttcaa aaatgcctgg gctgacaacg caaatgcttg tgccaagcaa tatgcgggaa
     1981 ctggtgcctt gaagactgac tttaccagaa ctggaaagag aactcatttg ggacttataa
     2041 tggatggctg gaactcaatg atacgatatt ataagaacaa cttttccgat ggatttagac
     2101 aagattccat agacttattt cttggaaact attcagtgga tgaattagaa tctcatagtc
     2161 ctttaagtgt tccaagggac tggaaattgt cgacggtacc gatgggagtg caggtggaaa
     2221 ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact
     2281 acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga aacaagccct
     2341 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     2401 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     2461 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     2521 tggaacggga tccaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca
     2581 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     2641 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     2701 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     2761 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     2821 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     2881 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     2941 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     3001 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     3061 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     3121 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     3181 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     3241 acgagctgta caagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3301 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3361 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3481 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3541 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3601 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3661 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3721 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3781 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3841 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3901 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3961 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttt aataatattg
     4081 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4201 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4261 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4321 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4381 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4441 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4501 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4561 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4621 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4681 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4741 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4801 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4861 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4921 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4981 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5041 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5101 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5161 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5221 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5281 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5341 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5401 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5461 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5521 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5581 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5641 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5701 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5761 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5821 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5881 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5941 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6001 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6061 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6121 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6181 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6241 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6301 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6361 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6421 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6481 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6541 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcca
//
