LOCUS       sequence-207728-        7029 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 7029)
  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 7029)
  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-HLx10
FEATURES             Location/Qualifiers
     source          1..7029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..317
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        318..521
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     472..492
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      560..569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             566..1273
                     /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(711..729)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     979..998
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1334..1654
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3770..3891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3807..3826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3861..3880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3898..4353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4247..4266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4380..4484
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4486..4843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4486..4677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4509..4529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4694..4829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4878..5672
                     /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             4878..5672
                     /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(4932..4951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5542..5561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5860..5879)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5904..5951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6280..6868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6769..6788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
       61 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      301 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      361 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      421 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      481 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      541 tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat
      601 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
      661 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      721 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
      781 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
      841 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
      901 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
      961 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     1021 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     1081 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     1141 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     1201 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     1261 cgagctgtac aagtccggac tcagatctcg aagcgcggcc gcgggagcag gaggagcagc
     1321 tcgagcggcg ctgggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc
     1381 caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg gaaagaaatt
     1441 tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat
     1501 ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat
     1561 atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac
     1621 tctcgtcttc gatgtggagc ttctaaaact ggaaagtgct ggtggtagtg ctggtggtag
     1681 tgctggtggt agtgctggtg gtagtgctgg tggtccgcga gctcaagctt cgaattctgc
     1741 aatggcgacg gcggcctacg agcagctgaa gctgcatatc acacctgaaa aattttatgt
     1801 ggaagcttgt gatgatggag cagatgacgt acttaccatt gaccgtgtgt ccacagaggt
     1861 tacccttgca gtcaagaaag atgttcctcc ttcagctgtc acaagaccaa tatttggtat
     1921 actgggcaca atccatctgg tggcaggtaa ttatcttata gtcattacca aaaagataaa
     1981 agtaggtgaa tttttcagtc atgtagtctg gaaagcaaca gattttgatg tcctttctta
     2041 taagaagaca atgttgcact taactgatat tcagttacaa gataataaaa ccttcctagc
     2101 gatgctaaac catgtcttga atgtggatgg attttacttt tcaacaacat atgatttgac
     2161 ccatactttg cagcggctat ccaacactag tcctgaattc caagaaatga gtctcttgga
     2221 aagggcagat cagcggtttg tatggaatgg tcatcttcta agagaacttt ctgcacagcc
     2281 agaggttcat cggtttgccc ttccagtgtt acatggcttt attaccatgc attcatgttc
     2341 tattaatgga aaatactttg attggattct catctcgagg aggagctgtt tcagagctgg
     2401 tgtgcgctat tatgtaagag gaattgattc ggaaggccat gcagctaact ttgtagaaac
     2461 agaacaaatt gtgcactaca atgggagcaa agcttcgttt gtacagactc gaggatcaat
     2521 acctgttttc tggtcccaaa gaccaaacct caagtacaaa ccactgccac agatcagcaa
     2581 agtagcaaat cacatggacg gtttccaaag gcattttgat tcccaagtaa ttatttatgg
     2641 aaaacaagtt ataatcaatc tgattaacca gaagggctcg gagaagccac ttgagcagac
     2701 atttgcaaca atggtgtctt ccttgggaag tggaatgatg agatacattg cctttgactt
     2761 ccataaggaa tgtaaaaata tgagatggga tcgactaagt attttattgg atcaggtagc
     2821 agaaatgcaa gatgaattaa gttattttct agtggactct gctggccagg tggtggcaaa
     2881 ccaggaaggc gtgttccgaa gcaattgcat ggattgtcta gatagaacca atgtgatcca
     2941 gagtttgtta gctcgtcgtt cacttcaggc ccaacttcag agactaggag ttttgcatgt
     3001 gggacaaaag cttgaagaac aagatgaatt tgagaagatt ttcaaaaatg cctgggctga
     3061 caacgcaaat gcttgtgcca agcaatatgc gggaactggt gccttgaaga ctgactttac
     3121 cagaactgga aagagaactc atttgggact tataatggat ggctggaact caatgatacg
     3181 atattataag aacaactttt ccgatggatt tagacaagat tccatagact tatttcttgg
     3241 aaactattca gtggatgaat tagaatctca tagtccttta agtgttccaa gggactggaa
     3301 agaagccgct gctagagaag ctgccgcaag agaggccgca gccagagagg ctgctgccag
     3361 ggaagcagca gctcgggaag ccgcagcacg cgaggctgca gcaagagaag cagccgcacg
     3421 cgaagcagcc gctagagagg cagctgctag attcctggct ttgcctatta tcatggttgt
     3481 tgccttttca atgtgcatta tctgtttgct tatggctggt gacacttgga cagaaacact
     3541 ggcctatgtg ctcttctggg gagttgcaag cattggaaca ttttttatca ttctttacaa
     3601 tggcaaagat tttgtcgatg ctcccagact ggtccagaaa gaaaagatag actgaagata
     3661 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3721 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     3901 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3961 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4021 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4081 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4141 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4201 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4261 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4321 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4501 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4561 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4621 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4681 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4741 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4801 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4861 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4921 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4981 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5041 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5101 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5161 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5221 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5281 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5341 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5401 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5461 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5521 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5581 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5641 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5701 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5761 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5821 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5881 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5941 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6001 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6061 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6121 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6181 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6421 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6901 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6961 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
     7021 atagcccat
//
