LOCUS       sequence-118630-        6395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express EGFP-tagged FKBP for heterodimerization with FRB in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6395)
  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 6395)
  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: pC4M-F2E-GFP-FKBP
FEATURES             Location/Qualifiers
     source          1..6395
                     /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"
     CDS             join(662..664,665..667,668..1378)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(662..664,665..667,668..1378)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(707..728)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(968..987)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1315..1336
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1385..1705
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1712..2032
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1712..2032
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2039..2065
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2097..2669
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2801..2820)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2866..2921
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2920..2939)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      3274..3409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3515..3532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3686..4274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3766..3785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5068..5087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5318..5422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5490..5508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5546..5568)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5668..5687
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5754..6209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5891..5912)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6103..6122
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6339..6361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6353..6370
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6354..6370
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6377..6395
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      721 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      781 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      841 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      901 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      961 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1021 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1081 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1141 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1201 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1261 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1321 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     1381 tagaggagtg caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg
     1441 ccagacctgc gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc
     1501 ccgggacaga aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg
     1561 ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga
     1621 ttatgcctat ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt
     1681 cgatgtggag cttctaaaac tggaaactag aggcgtgcag gtcgagacca tcagccccgg
     1741 cgacggccgc acctttccca agagaggcca gacttgcgtg gtccactaca ccggcatgct
     1801 ggaggacggc aagaagttcg acagcagccg cgaccgcaac aagcccttca agttcatgct
     1861 gggcaaacag gaagtgatcc gcggctggga ggaaggcgtg gctcagatga gcgtggggca
     1921 gcgggccaag ctgaccatca gccccgacta tgcctacggc gccaccggcc accccggcat
     1981 catccccccc cacgccaccc tggtgttcga cgtggagctg ctgaagctgg agactagtta
     2041 tccgtacgac gtaccagact acgcataaga aaagtgagga tcctgagaac ttcagggtga
     2101 gtttggggac ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag
     2161 ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac
     2221 cctcatgata attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta
     2281 ttttcttttc attttctgta actttttcgt taaactttag cttgcatttg taacgaattt
     2341 ttaaattcac ttttgtttat ttgtcagatt gtaagtactt tctctaatca cttttttttc
     2401 aaggcaatca gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa
     2461 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta
     2521 ttggtagaaa caactacacc ctggtcatca tcctgccttt ctctttatgg ttacaatgat
     2581 atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc
     2641 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt
     2701 ctcatcattt tggcaaagga ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     2761 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     2821 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     2881 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     2941 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca
     3001 tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc
     3061 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     3121 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     3181 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga
     3241 agatccctcg aggagctttt tgcaaaagcc ctaggcctcc aaaaaagcct cttcactact
     3301 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     3361 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     3421 ggggcgggac tatggttgct gactaattgc tgcattaatg aatcggccaa cgcgcgggga
     3481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5101 tgctcatcat tgccacctcc acctgaaaac gttcttcggg gcgaaaactc tcaaggatct
     5161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5461 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5521 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5581 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5641 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5701 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     5761 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5821 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5881 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5941 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6001 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6061 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6121 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6181 cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg ccattcaggc tgcgcaactg
     6241 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6301 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     6361 gacggccagt gaattgtaat acgactcact atagg
//
