LOCUS       sequence-207723-        6999 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 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-SAC1-HLx8
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        75..378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        379..582
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     533..553
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      621..630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             627..1334
                     /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(772..790)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1040..1059
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1395..1715
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3801..3922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3838..3857)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3892..3911
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3929..4384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4066..4087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4278..4297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4411..4515
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4517..4874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4517..4708
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4540..4560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4725..4860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4909..5703
                     /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             4909..5703
                     /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(4963..4982)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5573..5592
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5891..5910)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5935..5982
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6311..6899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6800..6819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
       61 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      121 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      181 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      241 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      301 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      361 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      421 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      481 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      541 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      601 atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca
      661 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      721 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      781 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      841 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      901 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      961 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1021 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1081 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1141 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1201 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1261 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1321 acgagctgta caagtccgga ctcagatctc gaagcgcggc cgcgggagca ggaggagcag
     1381 ctcgagcggc gctgggagtg caggtggaaa ccatctcccc aggagacggg cgcaccttcc
     1441 ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat ggaaagaaat
     1501 ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag caggaggtga
     1561 tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc aaactgacta
     1621 tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca ccacatgcca
     1681 ctctcgtctt cgatgtggag cttctaaaac tggaaagtgc tggtggtagt gctggtggta
     1741 gtgctggtgg tagtgctggt ggtagtgctg gtggtccgcg agctcaagct tcgaattctg
     1801 caatggcgac ggcggcctac gagcagctga agctgcatat cacacctgaa aaattttatg
     1861 tggaagcttg tgatgatgga gcagatgacg tacttaccat tgaccgtgtg tccacagagg
     1921 ttacccttgc agtcaagaaa gatgttcctc cttcagctgt cacaagacca atatttggta
     1981 tactgggcac aatccatctg gtggcaggta attatcttat agtcattacc aaaaagataa
     2041 aagtaggtga atttttcagt catgtagtct ggaaagcaac agattttgat gtcctttctt
     2101 ataagaagac aatgttgcac ttaactgata ttcagttaca agataataaa accttcctag
     2161 cgatgctaaa ccatgtcttg aatgtggatg gattttactt ttcaacaaca tatgatttga
     2221 cccatacttt gcagcggcta tccaacacta gtcctgaatt ccaagaaatg agtctcttgg
     2281 aaagggcaga tcagcggttt gtatggaatg gtcatcttct aagagaactt tctgcacagc
     2341 cagaggttca tcggtttgcc cttccagtgt tacatggctt tattaccatg cattcatgtt
     2401 ctattaatgg aaaatacttt gattggattc tcatctcgag gaggagctgt ttcagagctg
     2461 gtgtgcgcta ttatgtaaga ggaattgatt cggaaggcca tgcagctaac tttgtagaaa
     2521 cagaacaaat tgtgcactac aatgggagca aagcttcgtt tgtacagact cgaggatcaa
     2581 tacctgtttt ctggtcccaa agaccaaacc tcaagtacaa accactgcca cagatcagca
     2641 aagtagcaaa tcacatggac ggtttccaaa ggcattttga ttcccaagta attatttatg
     2701 gaaaacaagt tataatcaat ctgattaacc agaagggctc ggagaagcca cttgagcaga
     2761 catttgcaac aatggtgtct tccttgggaa gtggaatgat gagatacatt gcctttgact
     2821 tccataagga atgtaaaaat atgagatggg atcgactaag tattttattg gatcaggtag
     2881 cagaaatgca agatgaatta agttattttc tagtggactc tgctggccag gtggtggcaa
     2941 accaggaagg cgtgttccga agcaattgca tggattgtct agatagaacc aatgtgatcc
     3001 agagtttgtt agctcgtcgt tcacttcagg cccaacttca gagactagga gttttgcatg
     3061 tgggacaaaa gcttgaagaa caagatgaat ttgagaagat tttcaaaaat gcctgggctg
     3121 acaacgcaaa tgcttgtgcc aagcaatatg cgggaactgg tgccttgaag actgacttta
     3181 ccagaactgg aaagagaact catttgggac ttataatgga tggctggaac tcaatgatac
     3241 gatattataa gaacaacttt tccgatggat ttagacaaga ttccatagac ttatttcttg
     3301 gaaactattc agtggatgaa ttagaatctc atagtccttt aagtgttcca agggactgga
     3361 aagaagccgc tgctagagaa gctgccgcaa gagaggccgc agccagagag gctgctgcca
     3421 gggaagcagc agctcgggaa gccgcagcac gcgaggctgc agcaagagaa gcagccgcac
     3481 gcttcctggc tttgcctatt atcatggttg ttgccttttc aatgtgcatt atctgtttgc
     3541 ttatggctgg tgacacttgg acagaaacac tggcctatgt gctcttctgg ggagttgcaa
     3601 gcattggaac attttttatc attctttaca atggcaaaga ttttgtcgat gctcccagac
     3661 tggtccagaa agaaaagata gactgaagat aactgatcat aatcagccat accacatttg
     3721 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3781 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3841 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3901 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     3961 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4021 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4081 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4141 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4201 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4261 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4321 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4381 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4501 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4561 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4621 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4681 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4801 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4861 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4921 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4981 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5041 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     5101 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5161 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5221 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5281 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5341 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5401 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     5461 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5521 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5581 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5641 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5701 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5761 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5821 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5881 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5941 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     6001 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     6061 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     6121 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     6181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6961 tctttcctgc gttatcccct gattctgtgg ataaccgta
//
