LOCUS       sequence-118629-        5473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express Fis1 tail-anchored FRB for heterodimerization with FKBP in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5473)
  AUTHORS   Narendra D, Kane LA, Hauser DN, Fearnley IM, Youle RJ
  TITLE     p62/SQSTM1 is required for Parkin-induced mitochondrial clustering 
            but not mitophagy; VDAC1 is dispensable for both.
  JOURNAL   Autophagy. 2010 Nov;6(8):1090-106.
  PUBMED    20890124
REFERENCE   2  (bases 1 to 5473)
  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: 
            "Autophagy."; date: "2010-11"; volume: "6(8)"; pages: "1090-106"
COMMENT     Sequence Label: pC4-RhE-FRB-Fis1
FEATURES             Location/Qualifiers
     source          1..5473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        323..526
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     477..497
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     523..547
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     5'UTR           616..655
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             688..966
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     intron          1175..1747
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1879..1898)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1944..1999
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1998..2017)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      2352..2487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2593..2610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2764..3352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2844..2863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4146..4165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4396..4500)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4568..4586
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4624..4646)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4746..4765
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4832..5287)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4969..4990)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5181..5200
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5417..5439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5431..5448
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5432..5448
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5455..5473
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgcgttcga gctcgccccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
       61 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      181 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      241 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      301 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      421 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      481 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
      541 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      601 atccagcctc cgggggatct tggtggcgtg aaactcccgc acctcttcgg ccagcgaatt
      661 ccagaagccg ccaccatggc ttctagaatc ctctggcatg agatgtggca tgaaggcctg
      721 gaagaggcat ctcgtttgta ctttggggaa aggaacgtga aaggcatgtt tgaggtgctg
      781 gagcccttgc atgctatgat ggaacggggc ccccagactc tgaaggaaac atcctttaat
      841 caggcctatg gtcgagattt aatggaggcc caagagtggt gcaggaagta catgaaatca
      901 gggaatgtca aggacctcct ccaagcctgg gacctctatt atcatgtgtt ccgacgaatc
      961 tcaaagacta gttacgtccg cgggttgctg cagacagagc cccagaacaa ccaggccaag
     1021 gaactggagc ggctcattga caaggccatg aagaaagatg gactcgtggg catggccatc
     1081 gtgggaggca tggccctggg tgtggcggga ctggccggac tcatcggact tgctgtgtcc
     1141 aagtccaaat cctgaggatc ctgagaactt cagggtgagt ttggggaccc ttgattgttc
     1201 tttctttttc gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt
     1261 gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt
     1321 tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac
     1381 tttttcgtta aactttagct tgcatttgta acgaattttt aaattcactt ttgtttattt
     1441 gtcagattgt aagtactttc tctaatcact tttttttcaa ggcaatcagg gtatattata
     1501 ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt
     1561 tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacatcct
     1621 ggtcatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat
     1681 aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttcttttt
     1741 cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaaggatt
     1801 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1861 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     1921 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     1981 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     2041 gaatgagtat ttggtttaga gtttggcaac atatgccata tgctggctgc catgaacaaa
     2101 ggtggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     2161 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     2221 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     2281 gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgag gagctttttg
     2341 caaaagccct aggcctccaa aaaagcctct tcactacttc tggaatagct cagaggccga
     2401 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     2461 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     2521 ctaattgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2581 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2641 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2701 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3181 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     3241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3841 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3901 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3961 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4021 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4081 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4141 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg ccacctccac
     4201 ctgaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4501 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4561 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4621 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4681 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4741 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4801 cagatgcgta aggagaaaat accgcatcag gaaattgtaa acgttaatat tttgttaaaa
     4861 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4921 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4981 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5041 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5101 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5161 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5221 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5281 ggcgcgtcgc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5341 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5401 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     5461 gactcactat agg
//
