LOCUS       sequence-207717-        6909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable SAC1 with extended linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6909)
  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 6909)
  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-SAC1-HLx2
FEATURES             Location/Qualifiers
     source          1..6909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        352..555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     506..526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      594..603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             600..1307
                     /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(745..763)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1013..1032
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1368..1688
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3684..3805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3721..3740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3775..3794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3812..4267)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3949..3970)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4161..4180
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4294..4398
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4400..4757
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4400..4591
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4423..4443)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4608..4743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4792..5586
                     /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             4792..5586
                     /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(4846..4865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5456..5475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5774..5793)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5818..5865
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6194..6782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6683..6702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
       61 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      121 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      181 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      241 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      301 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      361 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      421 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      481 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      541 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca
      601 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      661 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      721 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      781 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      841 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      901 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
      961 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     1021 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1081 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1141 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1201 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1261 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc ggactcagat
     1321 ctcgaagcgc ggccgcggga gcaggaggag cagctcgagc ggcgctggga gtgcaggtgg
     1381 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
     1441 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
     1501 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
     1561 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
     1621 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
     1681 aactggaaag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg
     1741 ctggtggtcc gcgagctcaa gcttcgaatt ctgcaatggc gacggcggcc tacgagcagc
     1801 tgaagctgca tatcacacct gaaaaatttt atgtggaagc ttgtgatgat ggagcagatg
     1861 acgtacttac cattgaccgt gtgtccacag aggttaccct tgcagtcaag aaagatgttc
     1921 ctccttcagc tgtcacaaga ccaatatttg gtatactggg cacaatccat ctggtggcag
     1981 gtaattatct tatagtcatt accaaaaaga taaaagtagg tgaatttttc agtcatgtag
     2041 tctggaaagc aacagatttt gatgtccttt cttataagaa gacaatgttg cacttaactg
     2101 atattcagtt acaagataat aaaaccttcc tagcgatgct aaaccatgtc ttgaatgtgg
     2161 atggatttta cttttcaaca acatatgatt tgacccatac tttgcagcgg ctatccaaca
     2221 ctagtcctga attccaagaa atgagtctct tggaaagggc agatcagcgg tttgtatgga
     2281 atggtcatct tctaagagaa ctttctgcac agccagaggt tcatcggttt gcccttccag
     2341 tgttacatgg ctttattacc atgcattcat gttctattaa tggaaaatac tttgattgga
     2401 ttctcatctc gaggaggagc tgtttcagag ctggtgtgcg ctattatgta agaggaattg
     2461 attcggaagg ccatgcagct aactttgtag aaacagaaca aattgtgcac tacaatggga
     2521 gcaaagcttc gtttgtacag actcgaggat caatacctgt tttctggtcc caaagaccaa
     2581 acctcaagta caaaccactg ccacagatca gcaaagtagc aaatcacatg gacggtttcc
     2641 aaaggcattt tgattcccaa gtaattattt atggaaaaca agttataatc aatctgatta
     2701 accagaaggg ctcggagaag ccacttgagc agacatttgc aacaatggtg tcttccttgg
     2761 gaagtggaat gatgagatac attgcctttg acttccataa ggaatgtaaa aatatgagat
     2821 gggatcgact aagtatttta ttggatcagg tagcagaaat gcaagatgaa ttaagttatt
     2881 ttctagtgga ctctgctggc caggtggtgg caaaccagga aggcgtgttc cgaagcaatt
     2941 gcatggattg tctagataga accaatgtga tccagagttt gttagctcgt cgttcacttc
     3001 aggcccaact tcagagacta ggagttttgc atgtgggaca aaagcttgaa gaacaagatg
     3061 aatttgagaa gattttcaaa aatgcctggg ctgacaacgc aaatgcttgt gccaagcaat
     3121 atgcgggaac tggtgccttg aagactgact ttaccagaac tggaaagaga actcatttgg
     3181 gacttataat ggatggctgg aactcaatga tacgatatta taagaacaac ttttccgatg
     3241 gatttagaca agattccata gacttatttc ttggaaacta ttcagtggat gaattagaat
     3301 ctcatagtcc tttaagtgtt ccaagggact ggaaagaagc cgctgctaga gaagctgccg
     3361 caagattcct ggctttgcct attatcatgg ttgttgcctt ttcaatgtgc attatctgtt
     3421 tgcttatggc tggtgacact tggacagaaa cactggccta tgtgctcttc tggggagttg
     3481 caagcattgg aacatttttt atcattcttt acaatggcaa agattttgtc gatgctccca
     3541 gactggtcca gaaagaaaag atagactgaa gataactgat cataatcagc cataccacat
     3601 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3661 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3721 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3781 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     3841 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3901 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3961 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4021 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4081 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4141 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4201 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4261 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4321 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4381 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4441 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4501 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4561 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4621 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4681 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4741 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4801 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4861 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4921 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4981 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5041 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5101 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5161 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5221 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5281 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5341 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5401 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5461 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5521 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5581 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5641 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5701 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5761 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5821 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5881 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5941 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6001 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6061 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6121 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6181 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6241 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6301 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6361 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6421 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6481 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6541 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6601 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6661 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6721 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6781 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6841 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     6901 tattaatag
//
