LOCUS       sequence-207718-        6939 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 6939)
  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 6939)
  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-HLx4
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        70..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        374..577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     528..548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      616..625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             622..1329
                     /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(767..785)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1035..1054
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1390..1710
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3736..3857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3773..3792)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3827..3846
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3864..4319)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4001..4022)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4213..4232
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4346..4450
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4452..4809
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4452..4643
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4475..4495)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4660..4795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4844..5638
                     /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             4844..5638
                     /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(4898..4917)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5508..5527
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5826..5845)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5870..5917
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6246..6834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6735..6754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
       61 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      121 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      181 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      241 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      301 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      361 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      421 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      481 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      541 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      601 ctagcgctac cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc
      661 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      721 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      781 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
      841 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
      901 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      961 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1021 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1081 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     1141 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1201 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     1261 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgaagc gcggccgcgg gagcaggagg agcagctcga
     1381 gcggcgctgg gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag
     1441 cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat
     1501 tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga
     1561 ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct
     1621 ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc
     1681 gtcttcgatg tggagcttct aaaactggaa agtgctggtg gtagtgctgg tggtagtgct
     1741 ggtggtagtg ctggtggtag tgctggtggt ccgcgagctc aagcttcgaa ttctgcaatg
     1801 gcgacggcgg cctacgagca gctgaagctg catatcacac ctgaaaaatt ttatgtggaa
     1861 gcttgtgatg atggagcaga tgacgtactt accattgacc gtgtgtccac agaggttacc
     1921 cttgcagtca agaaagatgt tcctccttca gctgtcacaa gaccaatatt tggtatactg
     1981 ggcacaatcc atctggtggc aggtaattat cttatagtca ttaccaaaaa gataaaagta
     2041 ggtgaatttt tcagtcatgt agtctggaaa gcaacagatt ttgatgtcct ttcttataag
     2101 aagacaatgt tgcacttaac tgatattcag ttacaagata ataaaacctt cctagcgatg
     2161 ctaaaccatg tcttgaatgt ggatggattt tacttttcaa caacatatga tttgacccat
     2221 actttgcagc ggctatccaa cactagtcct gaattccaag aaatgagtct cttggaaagg
     2281 gcagatcagc ggtttgtatg gaatggtcat cttctaagag aactttctgc acagccagag
     2341 gttcatcggt ttgcccttcc agtgttacat ggctttatta ccatgcattc atgttctatt
     2401 aatggaaaat actttgattg gattctcatc tcgaggagga gctgtttcag agctggtgtg
     2461 cgctattatg taagaggaat tgattcggaa ggccatgcag ctaactttgt agaaacagaa
     2521 caaattgtgc actacaatgg gagcaaagct tcgtttgtac agactcgagg atcaatacct
     2581 gttttctggt cccaaagacc aaacctcaag tacaaaccac tgccacagat cagcaaagta
     2641 gcaaatcaca tggacggttt ccaaaggcat tttgattccc aagtaattat ttatggaaaa
     2701 caagttataa tcaatctgat taaccagaag ggctcggaga agccacttga gcagacattt
     2761 gcaacaatgg tgtcttcctt gggaagtgga atgatgagat acattgcctt tgacttccat
     2821 aaggaatgta aaaatatgag atgggatcga ctaagtattt tattggatca ggtagcagaa
     2881 atgcaagatg aattaagtta ttttctagtg gactctgctg gccaggtggt ggcaaaccag
     2941 gaaggcgtgt tccgaagcaa ttgcatggat tgtctagata gaaccaatgt gatccagagt
     3001 ttgttagctc gtcgttcact tcaggcccaa cttcagagac taggagtttt gcatgtggga
     3061 caaaagcttg aagaacaaga tgaatttgag aagattttca aaaatgcctg ggctgacaac
     3121 gcaaatgctt gtgccaagca atatgcggga actggtgcct tgaagactga ctttaccaga
     3181 actggaaaga gaactcattt gggacttata atggatggct ggaactcaat gatacgatat
     3241 tataagaaca acttttccga tggatttaga caagattcca tagacttatt tcttggaaac
     3301 tattcagtgg atgaattaga atctcatagt cctttaagtg ttccaaggga ctggaaagaa
     3361 gccgctgcta gagaagctgc cgcaagagag gccgcagcca gagaggctgc tgccaggttc
     3421 ctggctttgc ctattatcat ggttgttgcc ttttcaatgt gcattatctg tttgcttatg
     3481 gctggtgaca cttggacaga aacactggcc tatgtgctct tctggggagt tgcaagcatt
     3541 ggaacatttt ttatcattct ttacaatggc aaagattttg tcgatgctcc cagactggtc
     3601 cagaaagaaa agatagactg aagataactg atcataatca gccataccac atttgtagag
     3661 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3721 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3781 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3841 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3901 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3961 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4021 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4081 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4141 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4201 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4261 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4441 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4561 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4621 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4681 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4741 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4801 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4861 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4921 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4981 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     5041 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5101 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5161 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5221 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5281 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5341 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     5401 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5461 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5521 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5581 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5641 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5701 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5761 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5821 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5881 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5941 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     6001 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     6061 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     6121 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6181 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6361 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6421 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6481 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6541 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6601 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6661 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6721 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6781 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6841 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6901 cctgcgttat cccctgattc tgtggataac cgtattacc
//
