LOCUS       sequence-207712-        6689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable cytosolic SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6689)
  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 6689)
  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-SAC1deltaTMD
FEATURES             Location/Qualifiers
     source          1..6689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        779..1082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1083..1286
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1237..1257
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1325..1334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1331..2038
                     /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(1476..1494)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1744..1763
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2099..2419
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    4195..4316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4232..4251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4286..4305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4323..4778)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4460..4481)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4672..4691
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4805..4909
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4911..5268
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4911..5102
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4934..4954)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5119..5254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5303..6097
                     /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             5303..6097
                     /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(5357..5376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5967..5986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6285..6304)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6329..6376
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
      721 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      781 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      841 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      901 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      961 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1021 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1081 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     1141 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1201 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1261 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     1321 ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     1381 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     1441 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     1501 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     1561 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     1621 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1681 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1741 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     1801 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     1861 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     1921 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1981 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc
     2041 cggactcaga tctcgaagcg cggccgcggg agcaggagga gcagctcgag cggcgctggg
     2101 agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac
     2161 ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaatttgatt cctcccggga
     2221 cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga
     2281 aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc
     2341 ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt
     2401 ggagcttcta aaactggaaa gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc
     2461 tggtggtagt gctggtggtc cgcgagctca agcttcgaat tctgcagagc tcatggcgac
     2521 ggcggcctac gagcagctga agctgcatat cacacctgaa aaattttatg tggaagcttg
     2581 tgatgatgga gcagatgacg tacttaccat tgaccgtgtg tccacagagg ttacccttgc
     2641 agtcaagaaa gatgttcctc cttcagctgt cacaagacca atatttggta tactgggcac
     2701 aatccatctg gtggcaggta attatcttat agtcattacc aaaaagataa aagtaggtga
     2761 atttttcagt catgtagtct ggaaagcaac agattttgat gtcctttctt ataagaagac
     2821 aatgttgcac ttaactgata ttcagttaca agataataaa accttcctag cgatgctaaa
     2881 ccatgtcttg aatgtggatg gattttactt ttcaacaaca tatgatttga cccatacttt
     2941 gcagcggcta tccaacacta gtcctgaatt ccaagaaatg agtctcttgg aaagggcaga
     3001 tcagcggttt gtatggaatg gtcatcttct aagagaactt tctgcacagc cagaggttca
     3061 tcggtttgcc cttccagtgt tacatggctt tattaccatg cattcatgtt ctattaatgg
     3121 aaaatacttt gattggattc tcatctcgag gaggagctgt ttcagagctg gtgtgcgcta
     3181 ttatgtaaga ggaattgatt cggaaggcca tgcagctaac tttgtagaaa cagaacaaat
     3241 tgtgcactac aatgggagca aagcttcgtt tgtacagact cgaggatcaa tacctgtttt
     3301 ctggtcccaa agaccaaacc tcaagtacaa accactgcca cagatcagca aagtagcaaa
     3361 tcacatggac ggtttccaaa ggcattttga ttcccaagta attatttatg gaaaacaagt
     3421 tataatcaat ctgattaacc agaagggctc ggagaagcca cttgagcaga catttgcaac
     3481 aatggtgtct tccttgggaa gtggaatgat gagatacatt gcctttgact tccataagga
     3541 atgtaaaaat atgagatggg atcgactaag tattttattg gatcaggtag cagaaatgca
     3601 agatgaatta agttattttc tagtggactc tgctggccag gtggtggcaa accaggaagg
     3661 cgtgttccga agcaattgca tggattgtct agatagaacc aatgtgatcc agagtttgtt
     3721 agctcgtcgt tcacttcagg cccaacttca gagactagga gttttgcatg tgggacaaaa
     3781 gcttgaagaa caagatgaat ttgagaagat tttcaaaaat gcctgggctg acaacgcaaa
     3841 tgcttgtgcc aagcaatatg cgggaactgg tgccttgaag actgacttta ccagaactgg
     3901 aaagagaact catttgggac ttataatgga tggctggaac tcaatgatac gatattataa
     3961 gaacaacttt tccgatggat ttagacaaga ttccatagac ttatttcttg gaaactattc
     4021 agtggatgaa ttagaatctc atagtccttt aagtgttcca agggactgga aattgtcgac
     4081 tttaaactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4141 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4201 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4261 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     4321 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4381 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4441 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4501 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4561 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4621 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4681 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4741 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4801 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4861 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4921 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4981 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5041 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5101 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5161 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5221 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     5281 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5341 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5401 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5461 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     5521 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5581 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5641 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5701 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5761 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5821 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5881 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5941 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6001 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6061 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     6121 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     6181 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6241 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     6301 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     6361 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     6421 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     6481 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     6541 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6601 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6661 ttccactgag cgtcagaccc cgtagaaaa
//
