LOCUS       sequence-207652-        6586 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Recruitable cytosolic SAC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  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 6586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: SAC1deltaTMD-FKBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2208..2210,2211..2531)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      2547..2556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2553..3263
                     /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(2698..2716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2966..2985
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3385..3506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3422..3441)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3476..3495
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3513..3968)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3650..3671)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3862..3881
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3995..4099
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4101..4458
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4101..4292
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4124..4144)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4309..4444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4493..5287
                     /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             4493..5287
                     /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(4547..4566)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5157..5176
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5475..5494)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5519..5566
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5895..6483
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6384..6403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctcgagc tcatggcgac ggcggcctac gagcagctga
      661 agctgcatat cacacctgaa aaattttatg tggaagcttg tgatgatgga gcagatgacg
      721 tacttaccat tgaccgtgtg tccacagagg ttacccttgc agtcaagaaa gatgttcctc
      781 cttcagctgt cacaagacca atatttggta tactgggcac aatccatctg gtggcaggta
      841 attatcttat agtcattacc aaaaagataa aagtaggtga atttttcagt catgtagtct
      901 ggaaagcaac agattttgat gtcctttctt ataagaagac aatgttgcac ttaactgata
      961 ttcagttaca agataataaa accttcctag cgatgctaaa ccatgtcttg aatgtggatg
     1021 gattttactt ttcaacaaca tatgatttga cccatacttt gcagcggcta tccaacacta
     1081 gtcctgaatt ccaagaaatg agtctcttgg aaagggcaga tcagcggttt gtatggaatg
     1141 gtcatcttct aagagaactt tctgcacagc cagaggttca tcggtttgcc cttccagtgt
     1201 tacatggctt tattaccatg cattcatgtt ctattaatgg aaaatacttt gattggattc
     1261 tcatctcgag gaggagctgt ttcagagctg gtgtgcgcta ttatgtaaga ggaattgatt
     1321 cggaaggcca tgcagctaac tttgtagaaa cagaacaaat tgtgcactac aatgggagca
     1381 aagcttcgtt tgtacagact cgaggatcaa tacctgtttt ctggtcccaa agaccaaacc
     1441 tcaagtacaa accactgcca cagatcagca aagtagcaaa tcacatggac ggtttccaaa
     1501 ggcattttga ttcccaagta attatttatg gaaaacaagt tataatcaat ctgattaacc
     1561 agaagggctc ggagaagcca cttgagcaga catttgcaac aatggtgtct tccttgggaa
     1621 gtggaatgat gagatacatt gcctttgact tccataagga atgtaaaaat atgagatggg
     1681 atcgactaag tattttattg gatcaggtag cagaaatgca agatgaatta agttattttc
     1741 tagtggactc tgctggccag gtggtggcaa accaggaagg cgtgttccga agcaattgca
     1801 tggattgtct agatagaacc aatgtgatcc agagtttgtt agctcgtcgt tcacttcagg
     1861 cccaacttca gagactagga gttttgcatg tgggacaaaa gcttgaagaa caagatgaat
     1921 ttgagaagat tttcaaaaat gcctgggctg acaacgcaaa tgcttgtgcc aagcaatatg
     1981 cgggaactgg tgccttgaag actgacttta ccagaactgg aaagagaact catttgggac
     2041 ttataatgga tggctggaac tcaatgatac gatattataa gaacaacttt tccgatggat
     2101 ttagacaaga ttccatagac ttatttcttg gaaactattc agtggatgaa ttagaatctc
     2161 atagtccttt aagtgttcca agggactgga aattgtcgac ggtaccgatg ggagtgcagg
     2221 tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg
     2281 tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca
     2341 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     2401 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     2461 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     2521 taaaactgga acgggatcca ccggtcgcca ccatggtgag caagggcgag gaggataaca
     2581 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     2641 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2701 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     2761 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     2821 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2881 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     2941 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     3001 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     3061 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     3121 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     3181 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     3241 gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag ccataccaca
     3301 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3361 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3421 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3481 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3541 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3601 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3661 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3721 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3781 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3841 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3901 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3961 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgctttaata
     4081 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4141 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4201 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4321 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4381 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4441 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4921 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5041 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5281 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5341 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5401 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5461 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5521 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5581 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5641 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5701 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5761 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5821 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5881 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5941 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6001 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6061 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6121 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6181 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6241 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6301 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6361 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6421 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6481 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6541 atgttctttc ctgcgttatc ccctgattct gtggataacc gtatta
//
