LOCUS       sequence-203937-        6792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6792)
  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 6792)
  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-SAC1-FKBP
FEATURES             Location/Qualifiers
     source          1..6792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3136..3453
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="VQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPF
                     KFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLK
                     LE"
     polyA_signal    3580..3701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3617..3636)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3671..3690
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3708..4163)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3845..3866)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4057..4076
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4190..4294
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4296..4653
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4296..4487
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4319..4339)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4504..4639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4688..5482
                     /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             4688..5482
                     /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(4742..4761)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5352..5371
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5670..5689)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5714..5761
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6090..6678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6579..6598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctatggc gacggcggcc tacgagcagc tgaagctgca tatcacacct
     1381 gaaaaatttt atgtggaagc ttgtgatgat ggagcagatg acgtacttac cattgaccgt
     1441 gtgtccacag aggttaccct tgcagtcaag aaagatgttc ctccttcagc tgtcacaaga
     1501 ccaatatttg gtatactggg cacaatccat ctggtggcag gtaattatct tatagtcatt
     1561 accaaaaaga taaaagtagg tgaatttttc agtcatgtag tctggaaagc aacagatttt
     1621 gatgtccttt cttataagaa gacaatgttg cacttaactg atattcagtt acaagataat
     1681 aaaaccttcc tagcgatgct aaaccatgtc ttgaatgtgg atggatttta cttttcaaca
     1741 acatatgatt tgacccatac tttgcagcgg ctatccaaca ctagtcctga attccaagaa
     1801 atgagtctct tggaaagggc agatcagcgg tttgtatgga atggtcatct tctaagagaa
     1861 ctttctgcac agccagaggt tcatcggttt gcccttccag tgttacatgg ctttattacc
     1921 atgcattcat gttctattaa tggaaaatac tttgattgga ttctcatctc gaggaggagc
     1981 tgtttcagag ctggtgtgcg ctattatgta agaggaattg attcggaagg ccatgcagct
     2041 aactttgtag aaacagaaca aattgtgcac tacaatggga gcaaagcttc gtttgtacag
     2101 actcgaggat caatacctgt tttctggtcc caaagaccaa acctcaagta caaaccactg
     2161 ccacagatca gcaaagtagc aaatcacatg gacggtttcc aaaggcattt tgattcccaa
     2221 gtaattattt atggaaaaca agttataatc aatctgatta accagaaggg ctcggagaag
     2281 ccacttgagc agacatttgc aacaatggtg tcttccttgg gaagtggaat gatgagatac
     2341 attgcctttg acttccataa ggaatgtaaa aatatgagat gggatcgact aagtatttta
     2401 ttggatcagg tagcagaaat gcaagatgaa ttaagttatt ttctagtgga ctctgctggc
     2461 caggtggtgg caaaccagga aggcgtgttc cgaagcaatt gcatggattg tctagataga
     2521 accaatgtga tccagagttt gttagctcgt cgttcacttc aggcccaact tcagagacta
     2581 ggagttttgc atgtgggaca aaagcttgaa gaacaagatg aatttgagaa gatttacaaa
     2641 aatgcctggg ctgacaacgc aaatgcttgt gccaagcaat atgcgggaac tggtgccttg
     2701 aagactgact ttaccagaac tggaaagaga actcatttgg gacttataat ggatggctgg
     2761 aactcaatga tacgatatta taagaacaac ttttccgatg gatttagaca agattccata
     2821 gacttatttc ttggaaacta ttcagtggat gaattagaat ctcatagtcc tttaagtgtt
     2881 ccaagggact ggaaattcct ggctttgcct attatcatgg ttgttgcctt ttcaatgtgc
     2941 attatctgtt tgcttatggc tggtgacact tggacagaaa cactggccta tgtgctcttc
     3001 tggggagttg caagcattgg aacatttttt atcattcttt acaatggcaa agattttgtc
     3061 gatgctccca gactggtcca gaaagaaaag atagacggaa tttgtatttg tggaaagcgg
     3121 cttggcttgg gtaccgtgca ggtggaaacc atctccccag gagacgggcg caccttcccc
     3181 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaattt
     3241 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     3301 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     3361 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     3421 ctcgtcttcg atgtggagct tctaaaactg gaatccaccg gatctagata actgatcata
     3481 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3541 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3601 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3661 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     3721 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3781 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3841 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3901 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3961 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4021 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4081 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4141 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4201 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4261 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4321 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4381 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4501 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4561 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4621 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4681 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4741 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4801 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4861 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4921 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4981 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5041 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5101 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5161 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5221 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5281 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5341 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5401 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5461 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5521 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5581 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5641 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5701 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5761 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5821 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5881 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5941 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6001 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6061 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     6241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6721 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6781 accgccatgc at
//
