LOCUS       sequence-207685-        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        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      602..611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             608..1315
                     /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(753..771)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1021..1040
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3131..3448
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="VQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPF
                     KFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLK
                     LE"
     polyA_signal    3575..3696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3612..3631)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3666..3685
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3703..4158)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3840..3861)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4052..4071
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4185..4289
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4291..4648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4291..4482
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4314..4334)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4499..4634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4683..5477
                     /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             4683..5477
                     /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(4737..4756)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5347..5366
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5665..5684)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5709..5756
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6085..6673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6574..6593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt
      601 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
      661 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
      721 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
      781 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
      841 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
      901 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
      961 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1021 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     1081 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     1141 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1201 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1261 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtccgg
     1321 actcagatct atggcgacgg cggcctacga gcagctgaag ctgcatatca cacctgaaaa
     1381 attttatgtg gaagcttgtg atgatggagc agatgacgta cttaccattg accgtgtgtc
     1441 cacagaggtt acccttgcag tcaagaaaga tgttcctcct tcagctgtca caagaccaat
     1501 atttggtata ctgggcacaa tccatctggt ggcaggtaat tatcttatag tcattaccaa
     1561 aaagataaaa gtaggtgaat ttttcagtca tgtagtctgg aaagcaacag attttgatgt
     1621 cctttcttat aagaagacaa tgttgcactt aactgatatt cagttacaag ataataaaac
     1681 cttcctagcg atgctaaacc atgtcttgaa tgtggatgga ttttactttt caacaacata
     1741 tgatttgacc catactttgc agcggctatc caacactagt cctgaattcc aagaaatgag
     1801 tctcttggaa agggcagatc agcggtttgt atggaatggt catcttctaa gagaactttc
     1861 tgcacagcca gaggttcatc ggtttgccct tccagtgtta catggcttta ttaccatgca
     1921 ttcatgttct attaatggaa aatactttga ttggattctc atctcgagga ggagctgttt
     1981 cagagctggt gtgcgctatt atgtaagagg aattgattcg gaaggccatg cagctaactt
     2041 tgtagaaaca gaacaaattg tgcactacaa tgggagcaaa gcttcgtttg tacagactcg
     2101 aggatcaata cctgttttct ggtcccaaag accaaacctc aagtacaaac cactgccaca
     2161 gatcagcaaa gtagcaaatc acatggacgg tttccaaagg cattttgatt cccaagtaat
     2221 tatttatgga aaacaagtta taatcaatct gattaaccag aagggctcgg agaagccact
     2281 tgagcagaca tttgcaacaa tggtgtcttc cttgggaagt ggaatgatga gatacattgc
     2341 ctttgacttc cataaggaat gtaaaaatat gagatgggat cgactaagta ttttattgga
     2401 tcaggtagca gaaatgcaag atgaattaag ttattttcta gtggactctg ctggccaggt
     2461 ggtggcaaac caggaaggcg tgttccgaag caattgcatg gattgtctag atagaaccaa
     2521 tgtgatccag agtttgttag ctcgtcgttc acttcaggcc caacttcaga gactaggagt
     2581 tttgcatgtg ggacaaaagc ttgaagaaca agatgaattt gagaagattt tcaaaaatgc
     2641 ctgggctgac aacgcaaatg cttgtgccaa gcaatatgcg ggaactggtg ccttgaagac
     2701 tgactttacc agaactggaa agagaactca tttgggactt ataatggatg gctggaactc
     2761 aatgatacga tattataaga acaacttttc cgatggattt agacaagatt ccatagactt
     2821 atttcttgga aactattcag tggatgaatt agaatctcat agtcctttaa gtgttccaag
     2881 ggactggaaa ttcctggctt tgcctattat catggttgtt gccttttcaa tgtgcattat
     2941 ctgtttgctt atggctggtg acacttggac agaaacactg gcctatgtgc tcttctgggg
     3001 agttgcaagc attggaacat tttttatcat tctttacaat ggcaaagatt ttgtcgatgc
     3061 tcccagactg gtccagaaag aaaagataga cggaatttgt atttgtggaa agcggcttgg
     3121 cttgggtacc gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg
     3181 cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aatttgattc
     3241 ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg
     3301 ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc
     3361 agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt
     3421 cttcgatgtg gagcttctaa aactggaatc caccggatct agataactga tcataatcag
     3481 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3541 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3601 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3661 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3721 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3781 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3841 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3901 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3961 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4021 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4081 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4141 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4201 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4261 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4321 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4381 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4441 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4501 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4561 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4621 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4681 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4741 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4801 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4861 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4921 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4981 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5041 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5101 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5161 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5221 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5281 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5341 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5401 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5461 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5521 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5581 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5641 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5701 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5761 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5821 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5881 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5941 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6001 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6061 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6121 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6181 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6241 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6301 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6361 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6421 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6481 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6541 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6601 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6661 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6721 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6781 catgcattag tt
//
