LOCUS       sequence-395421-        7559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescently labeled membrane anchor C2 fused to the 
            dimerization domain FRB to recruit proteins to plasma membrane by 
            using the chemical dimerization system FKBP-FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  AUTHORS   Schelski M, Bradke F
  TITLE     Microtubule retrograde flow retains neuronal polarization in a 
            fluctuating state.
  JOURNAL   Sci Adv. 2022 Nov 4;8(44):eabo2336. doi: 10.1126/sciadv.abo2336. 
            Epub 2022 Nov 4.
  PUBMED    36332023
REFERENCE   2  (bases 1 to 7559)
  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: "Sci Adv."; 
            date: "2022-11-4"; volume: "8(44):eabo2336. doi"; pages: " 
            10.1126/sciadv.abo2336. Epub 2022 Nov 4"
COMMENT     Sequence Label: pBetaActin-dCer3-C2-FRB
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        587..862
                     /label=chicken beta-actin promoter
     primer_bind     1813..1835
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     complement(1938..1959)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2199..2218)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2546..2567
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3129..3407
                     /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"
     polyA_signal    3581..3715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3618..3637)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3672..3691
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3729..3745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3729..3745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3742..3764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3753..3769
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3777..3783,3784..3801,3802..3807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3822..3843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3960..3977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4131..4719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4211..4230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4890..5681,5682..5750))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5513..5532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5751..5855)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5923..5941
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5979..6001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6101..6120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6314..6336
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6328..6345
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6329..6345
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agtgcagcgc gcgttgcacc cggtgcccct tcccctccct tgcatcccag caggctgcac
       61 cccagcacca ggcccgtgca tgcatgctcc tggtgttatt gcagcctggt gcatgcatgc
      121 gtcttgatgg tgcagcgctg tgcatgcatc ctccttggtg tgtagcagtc tagtgcatgc
      181 atacccctcg gtgttattgc tgctctgtgc acgcacgctc attgtatcac ttcatcccag
      241 tgcatgcact cacactggag cgattgctgc tcggtgcacg cacactcatt gtatcactgc
      301 agctcagtgg ctgcacgcac accggtgtta ttgctgctcg gtgcgtgcat gcacatcagt
      361 gtcgctgcag ctcagtgcat gcacgctcat tgcccatcgc tatccctgcc tctcctgctg
      421 gcgctccccg ggaggtgact tcaaggggac cgcaggacca cctcgggggt ggggggaggg
      481 ctgcacacgc ggaccccgct ccccctcccc aacaaagcac tgtggaatca aaaagggggg
      541 aggggggatg gaggggcgcg tcacaccccc gccccacacc ctcacctcga ggtgagcccc
      601 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
      661 attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg
      721 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
      781 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
      841 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
      901 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      961 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggctcgt
     1021 ttcttttctg tggctgcgtg aaagccttaa agggctccgg gagggccctt tgtgcggggg
     1081 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
     1141 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     1201 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     1261 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcggcgg
     1321 tcgggctgta acccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     1381 ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg ntgccgggsg gggggkggcg
     1441 gcnaggkggg ggkgccgggc ggggncgggg ncncgcckcg ggccggggna gggnctncgg
     1501 gggrggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc
     1561 attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg
     1621 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt
     1681 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     1741 cccttctcca tctccagcct cggggctgtc cgcaggggga cggctgcctt cgggggggac
     1801 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ggntttatat cttcccttct
     1861 ctgttcctcc gcagccccca agctagctcg agatggtgag caagggcgag gagctgttca
     1921 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1981 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2041 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc tggggcgtgc
     2101 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2161 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2221 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2281 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacgcc atccacggca
     2341 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc ggcctcaact
     2401 gcaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2461 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2521 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccgggg
     2581 gaagttcagg aggttctagt tgcactgaac ccctaggcct gaaggataat accatcccca
     2641 acaagcagat cacagcctcc agctactaca aaacctgggg cctgagtgcc tttagctggt
     2701 ttccctacta cgcacgactg gataatcagg gcaagttcaa cgcctggacc gcccagacca
     2761 acagtgcctc tgagtggctg cagattgacc tgggctccca gaagcgagtc acgggcatca
     2821 tcacccaggg tgcccgagac tttggccaca ttcaatatgt ggctgcctac agggtggcct
     2881 atggtgatga tggtgtgacc tggactgagt acaaggaccc gggggcctca gaaagcaaga
     2941 ttttccctgg taacatggac aataattccc acaagaagaa catatttgag acgccgttcc
     3001 aggctcgctt cgtgcggatc cagcccgtgg cctggcacaa ccgtatcacc ctgcgagtgg
     3061 agctgctggg ctgttatgaa ttcagcatgg gggagtctgg tggatcaagt ggtggaatgg
     3121 cttctagaat cctctggcat gagatgtggc atgaaggcct ggaagaggca tctcgtttgt
     3181 actttgggga aaggaacgtg aaaggcatgt ttgaggtgct ggagcccttg catgctatga
     3241 tggaacgggg cccccagact ctgaaggaaa catcctttaa tcaggcctat ggtcgagatt
     3301 taatggaggc ccaagagtgg tgcaggaagt acatgaaatc agggaatgtc aaggacctcc
     3361 tccaagcctg ggacctctat tatcatgtgt tccgacgaat ctcaaagtaa aagcttcgaa
     3421 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataag taatgatcat
     3481 aatcagccat atcacatctg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3541 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3601 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3661 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaattc
     3721 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3781 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3841 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3901 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3961 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4021 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4201 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4261 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4321 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4381 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4441 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4501 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4561 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4621 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4681 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4741 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4801 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4861 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4921 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4981 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5041 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5101 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5161 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5221 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5281 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5341 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5401 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5461 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5521 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5581 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5641 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5701 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5761 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5821 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5881 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5941 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6001 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6061 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6121 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6181 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6241 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     6301 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gttgggatct
     6361 ttgcattggc ccacggctct caggatgggg atgctcccct tcagcacccg gttccccttg
     6421 gaaactgatg gtcctggctc tgtggcatgg cagtggcact gtgaggagcc ccctaccagc
     6481 agcacacagt ggggttggca ctgccacgct ccggatgccg cgctctgatc caaccccata
     6541 atcaagggaa cccgaattgc cccatcattg cccccaccac ccccatcctg ccgggccctc
     6601 acaccccacg ctgccttctg gtgacattcc ccagcccaaa cccacggctt catggctacc
     6661 gcggggcatt tcccattgcc gccccattat cagctctgca cacctcccgc tgtacccatg
     6721 cctcgtggct gcccttcttt gacgtataat cttctaatta atacccggcc ttgtcaaagt
     6781 ggagcacaaa cgttaattaa ttccccagca ggcaggtaat taacagtgtg actccctttt
     6841 tgctgcgagt ggggctgata cagagagatg tggcactatg gagcccacgg ggtcctggca
     6901 ctgggtgccc acggaggtcc ccatgtgctg cagtgtcacc gcctccgagg agacagtatt
     6961 gtccctgcgg tgtccctgca gctcagctct gtccacaggg ccacctccag tttggagggg
     7021 acacaatgca gccccgatgc aacccatcct cgcagcatcc cagggacaaa gaccccactg
     7081 caagaccgca cacagggctg ggtcccgctc ccctaatatc tacagtgctt cttgcatggc
     7141 cccttaatca atgcagttaa tcagcatgcg ctcatgcacc gctctggagc tgcaaagccc
     7201 ctcgcagcgc tgctcaccaa caccgcgcac cgccccggcc cagcctgcag cacgcgctgc
     7261 aaacaggaaa gaaacaaaat attgcccaaa tgtaggcaaa ggcattcggc tgccttgacc
     7321 tccgccgggc cgggccctgc ctgactcagc tccttactca gcgctcgctt cctccctccg
     7381 gctgccaccg ccgcagcgca caccctgaca aagagtggcc cttaacgggc tctgaggtgc
     7441 acccagcagt gcactcagca gtccaagggc cggcctggag gtttgcaccg ctacgtgctg
     7501 acattagcat tgaacttggc cctgggtagt gctgcaggcc gggcggggtg ggtgtagag
//
