LOCUS       sequence-207729-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable inactive SAC1 with extended linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  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 6999)
  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-HLx8
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             617..1324
                     /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(762..780)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1030..1049
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1385..1705
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3791..3912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3828..3847)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3882..3901
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3919..4374)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4056..4077)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4268..4287
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4401..4505
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4507..4864
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4507..4698
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4530..4550)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4715..4850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4899..5693
                     /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             4899..5693
                     /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(4953..4972)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5563..5582
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5881..5900)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5925..5972
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6301..6889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6790..6809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
      661 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      721 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      781 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
      841 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
      901 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
      961 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     1021 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     1081 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     1141 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     1201 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     1261 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     1321 caagtccgga ctcagatctc gaagcgcggc cgcgggagca ggaggagcag ctcgagcggc
     1381 gctgggagtg caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg
     1441 ccagacctgc gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc
     1501 ccgggacaga aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg
     1561 ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga
     1621 ttatgcctat ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt
     1681 cgatgtggag cttctaaaac tggaaagtgc tggtggtagt gctggtggta gtgctggtgg
     1741 tagtgctggt ggtagtgctg gtggtccgcg agctcaagct tcgaattctg caatggcgac
     1801 ggcggcctac gagcagctga agctgcatat cacacctgaa aaattttatg tggaagcttg
     1861 tgatgatgga gcagatgacg tacttaccat tgaccgtgtg tccacagagg ttacccttgc
     1921 agtcaagaaa gatgttcctc cttcagctgt cacaagacca atatttggta tactgggcac
     1981 aatccatctg gtggcaggta attatcttat agtcattacc aaaaagataa aagtaggtga
     2041 atttttcagt catgtagtct ggaaagcaac agattttgat gtcctttctt ataagaagac
     2101 aatgttgcac ttaactgata ttcagttaca agataataaa accttcctag cgatgctaaa
     2161 ccatgtcttg aatgtggatg gattttactt ttcaacaaca tatgatttga cccatacttt
     2221 gcagcggcta tccaacacta gtcctgaatt ccaagaaatg agtctcttgg aaagggcaga
     2281 tcagcggttt gtatggaatg gtcatcttct aagagaactt tctgcacagc cagaggttca
     2341 tcggtttgcc cttccagtgt tacatggctt tattaccatg cattcatgtt ctattaatgg
     2401 aaaatacttt gattggattc tcatctcgag gaggagctgt ttcagagctg gtgtgcgcta
     2461 ttatgtaaga ggaattgatt cggaaggcca tgcagctaac tttgtagaaa cagaacaaat
     2521 tgtgcactac aatgggagca aagcttcgtt tgtacagact cgaggatcaa tacctgtttt
     2581 ctggtcccaa agaccaaacc tcaagtacaa accactgcca cagatcagca aagtagcaaa
     2641 tcacatggac ggtttccaaa ggcattttga ttcccaagta attatttatg gaaaacaagt
     2701 tataatcaat ctgattaacc agaagggctc ggagaagcca cttgagcaga catttgcaac
     2761 aatggtgtct tccttgggaa gtggaatgat gagatacatt gcctttgact tccataagga
     2821 atgtaaaaat atgagatggg atcgactaag tattttattg gatcaggtag cagaaatgca
     2881 agatgaatta agttattttc tagtggactc tgctggccag gtggtggcaa accaggaagg
     2941 cgtgttccga agcaatagca tggattgtct agatagaacc aatgtgatcc agagtttgtt
     3001 agctcgtcgt tcacttcagg cccaacttca gagactagga gttttgcatg tgggacaaaa
     3061 gcttgaagaa caagatgaat ttgagaagat tttcaaaaat gcctgggctg acaacgcaaa
     3121 tgcttgtgcc aagcaatatg cgggaactgg tgccttgaag actgacttta ccagaactgg
     3181 aaagagaact catttgggac ttataatgga tggctggaac tcaatgatac gatattataa
     3241 gaacaacttt tccgatggat ttagacaaga ttccatagac ttatttcttg gaaactattc
     3301 agtggatgaa ttagaatctc atagtccttt aagtgttcca agggactgga aagaagccgc
     3361 tgctagagaa gctgccgcaa gagaggccgc agccagagag gctgctgcca gggaagcagc
     3421 agctcgggaa gccgcagcac gcgaggctgc agcaagagaa gcagccgcac gcttcctggc
     3481 tttgcctatt atcatggttg ttgccttttc aatgtgcatt atctgtttgc ttatggctgg
     3541 tgacacttgg acagaaacac tggcctatgt gctcttctgg ggagttgcaa gcattggaac
     3601 attttttatc attctttaca atggcaaaga ttttgtcgat gctcccagac tggtccagaa
     3661 agaaaagata gactgaagat aactgatcat aatcagccat accacatttg tagaggtttt
     3721 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3781 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3841 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3901 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3961 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4021 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4081 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4141 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4201 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4261 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4321 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4381 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4441 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4501 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4561 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4621 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4681 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4741 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4801 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4861 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4921 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4981 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5041 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5101 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5161 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5221 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5281 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5341 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5401 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5461 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5521 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5581 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5641 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5701 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5761 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5821 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5881 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5941 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6001 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6061 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6121 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6181 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6241 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6301 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6361 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6421 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6481 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6541 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6601 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6661 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6721 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6781 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6841 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6901 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6961 gttatcccct gattctgtgg ataaccgtat taccgccat
//
