LOCUS       sequence-207705-        6358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable inactive SAC1 (linker deletion).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6358)
  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 6358)
  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: SAC1delta452-587-FKBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6358
                     /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"
     CDS             1970..2290
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     regulatory      2306..2315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2312..3022
                     /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(2457..2475)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2725..2744
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3144..3265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3181..3200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3235..3254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3272..3727)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3409..3430)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3621..3640
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3754..3858
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3860..4217
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3860..4051
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3883..3903)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4068..4203
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4252..5046
                     /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             4252..5046
                     /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(4306..4325)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4916..4935
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5234..5253)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5278..5325
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5654..6242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6143..6162
                     /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 ggactcagat ctcgagctca tggcgacggc ggcctacgag cagctgaagc tgcatatcac
      661 acctgaaaaa ttttatgtgg aagcttgtga tgatggagca gatgacgtac ttaccattga
      721 ccgtgtgtcc acagaggtta cccttgcagt caagaaagat gttcctcctt cagctgtcac
      781 aagaccaata tttggtatac tgggcacaat ccatctggtg gcaggtaatt atcttatagt
      841 cattaccaaa aagataaaag taggtgaatt tttcagtcat gtagtctgga aagcaacaga
      901 ttttgatgtc ctttcttata agaagacaat gttgcactta actgatattc agttacaaga
      961 taataaaacc ttcctagcga tgctaaacca tgtcttgaat gtggatggat tttacttttc
     1021 aacaacatat gatttgaccc atactttgca gcggctatcc aacactagtc ctgaattcca
     1081 agaaatgagt ctcttggaaa gggcagatca gcggtttgta tggaatggtc atcttctaag
     1141 agaactttct gcacagccag aggttcatcg gtttgccctt ccagtgttac atggctttat
     1201 taccatgcat tcatgttcta ttaatggaaa atactttgat tggattctca tctcgaggag
     1261 gagctgtttc agagctggtg tgcgctatta tgtaagagga attgattcgg aaggccatgc
     1321 agctaacttt gtagaaacag aacaaattgt gcactacaat gggagcaaag cttcgtttgt
     1381 acagactcga ggatcaatac ctgttttctg gtcccaaaga ccaaacctca agtacaaacc
     1441 actgccacag atcagcaaag tagcaaatca catggacggt ttccaaaggc attttgattc
     1501 ccaagtaatt atttatggaa aacaagttat aatcaatctg attaaccaga agggctcgga
     1561 gaagccactt gagcagacat ttgcaacaat ggtgtcttcc ttgggaagtg gaatgatgag
     1621 atacattgcc tttgacttcc ataaggaatg taaaaatatg agatgggatc gactaagtat
     1681 tttattggat caggtagcag aaatgcaaga tgaattaagt tattttctag tggactctgc
     1741 tggccaggtg gtggcaaacc aggaaggcgt gttccgaagc aattgcatgg attgtctaga
     1801 tagaaccaat gtgatccaga gtttgttagc tcgtcgttca cttcaggccc aacttcagag
     1861 actaggagtt ttgcatgtgg gacaaaagct tgaagaacaa gatgaatttg agaagatttt
     1921 caaaaatgcc tgggctgaca acgcaaatgc ttgtgccaag caatatgcgg gagtgcaggt
     1981 ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt
     2041 gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg acagaaacaa
     2101 gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc
     2161 ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc
     2221 cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct
     2281 aaaactggaa cgggatccac cggtcgccac catggtgagc aagggcgagg aggataacat
     2341 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     2401 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2461 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2521 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2581 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2641 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2701 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2761 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2821 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2881 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2941 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     3001 catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc cataccacat
     3061 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3121 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3181 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3241 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3301 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3361 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3421 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3481 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3541 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3601 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3661 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3721 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3781 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gctttaataa
     3841 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3901 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3961 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4081 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4141 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4201 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4261 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4321 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4381 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4441 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4501 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4561 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4621 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4681 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4741 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4801 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4861 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4921 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4981 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5041 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5101 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5161 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5221 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5281 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5341 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5401 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5461 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5521 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5581 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5641 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5701 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5761 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5821 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5881 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5941 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6001 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6061 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6301 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcatta
//
