LOCUS       sequence-203936-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable inactive SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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-SAC1C389S
FEATURES             Location/Qualifiers
     source          1..6879
                     /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             1381..1701
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3667..3788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3704..3723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3758..3777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3795..4250)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3932..3953)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4144..4163
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4277..4381
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4383..4740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4383..4574
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4406..4426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4591..4726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4775..5569
                     /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             4775..5569
                     /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(4829..4848)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5439..5458
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5757..5776)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5801..5848
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6177..6765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6666..6685
                     /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 gatctcgaag cgcggccgcg ggagcaggag gagcagctcg agcggcgctg
     1381 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag
     1441 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
     1501 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
     1561 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
     1621 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
     1681 gtggagcttc taaaactgga aagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     1741 gctggtggta gtgctggtgg tccgcgagct caagcttcga attctgcaat ggcgacggcg
     1801 gcctacgagc agctgaagct gcatatcaca cctgaaaaat tttatgtgga agcttgtgat
     1861 gatggagcag atgacgtact taccattgac cgtgtgtcca cagaggttac ccttgcagtc
     1921 aagaaagatg ttcctccttc agctgtcaca agaccaatat ttggtatact gggcacaatc
     1981 catctggtgg caggtaatta tcttatagtc attaccaaaa agataaaagt aggtgaattt
     2041 ttcagtcatg tagtctggaa agcaacagat tttgatgtcc tttcttataa gaagacaatg
     2101 ttgcacttaa ctgatattca gttacaagat aataaaacct tcctagcgat gctaaaccat
     2161 gtcttgaatg tggatggatt ttacttttca acaacatatg atttgaccca tactttgcag
     2221 cggctatcca acactagtcc tgaattccaa gaaatgagtc tcttggaaag ggcagatcag
     2281 cggtttgtat ggaatggtca tcttctaaga gaactttctg cacagccaga ggttcatcgg
     2341 tttgcccttc cagtgttaca tggctttatt accatgcatt catgttctat taatggaaaa
     2401 tactttgatt ggattctcat ctcgaggagg agctgtttca gagctggtgt gcgctattat
     2461 gtaagaggaa ttgattcgga aggccatgca gctaactttg tagaaacaga acaaattgtg
     2521 cactacaatg ggagcaaagc ttcgtttgta cagactcgag gatcaatacc tgttttctgg
     2581 tcccaaagac caaacctcaa gtacaaacca ctgccacaga tcagcaaagt agcaaatcac
     2641 atggacggtt tccaaaggca ttttgattcc caagtaatta tttatggaaa acaagttata
     2701 atcaatctga ttaaccagaa gggctcggag aagccacttg agcagacatt tgcaacaatg
     2761 gtgtcttcct tgggaagtgg aatgatgaga tacattgcct ttgacttcca taaggaatgt
     2821 aaaaatatga gatgggatcg actaagtatt ttattggatc aggtagcaga aatgcaagat
     2881 gaattaagtt attttctagt ggactctgct ggccaggtgg tggcaaacca ggaaggcgtg
     2941 ttccgaagca atagcatgga ttgtctagat agaaccaatg tgatccagag tttgttagct
     3001 cgtcgttcac ttcaggccca acttcagaga ctaggagttt tgcatgtggg acaaaagctt
     3061 gaagaacaag atgaatttga gaagatttac aaaaatgcct gggctgacaa cgcaaatgct
     3121 tgtgccaagc aatatgcggg aactggtgcc ttgaagactg actttaccag aactggaaag
     3181 agaactcatt tgggacttat aatggatggc tggaactcaa tgatacgata ttataagaac
     3241 aacttttccg atggatttag acaagattcc atagacttat ttcttggaaa ctattcagtg
     3301 gatgaattag aatctcatag tcctttaagt gttccaaggg actggaaatt cctggctttg
     3361 cctattatca tggttgttgc cttttcaatg tgcattatct gtttgcttat ggctggtgac
     3421 acttggacag aaacactggc ctatgtgctc ttctggggag ttgcaagcat tggaacattt
     3481 tttatcattc tttacaatgg caaagatttt gtcgatgctc ccagactggt ccagaaagaa
     3541 aagatagact gaagataact gatcataatc agccatacca catttgtaga ggttttactt
     3601 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3661 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3781 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3841 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3901 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3961 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4021 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4081 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4141 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4201 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4261 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4321 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4381 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4561 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4621 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4681 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4741 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4801 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4861 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4921 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4981 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5041 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5101 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5161 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5221 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5281 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5341 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5401 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5461 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5521 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5581 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5641 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5701 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5761 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5821 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5881 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5941 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6001 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6301 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6841 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
