LOCUS       sequence-381708-        7095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescently labeled membrane anchor CAAX 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 7095)
  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 7095)
  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-FRB-dCer3-CAAX
FEATURES             Location/Qualifiers
     source          1..7095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    150..284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(187..206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     241..260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(298..314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(298..314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(311..333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    322..338
                     /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(346..352,353..370,371..376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    391..412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(529..546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(700..1288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(780..799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1459..2250,2251..2319))
                     /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     2082..2101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2320..2424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2492..2510
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2548..2570)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2670..2689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2883..2905
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2897..2914
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2898..2914
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4715..4990
                     /label=chicken beta-actin promoter
     primer_bind     5933..5955
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             6019..6297
                     /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"
     primer_bind     complement(6364..6385)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6625..6644)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6972..6993
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 acggtaccgc gggcccggga tccaccggat ctagataagt aatgatcata atcagccata
       61 tcacatctgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      121 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcg taatcatgtc
      301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1141 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2641 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2701 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     2761 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2821 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     2881 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ttgggatctt tgcattggcc
     2941 cacggctctc aggatgggga tgctcccctt cagcacccgg ttccccttgg aaactgatgg
     3001 tcctggctct gtggcatggc agtggcactg tgaggagccc cctaccagca gcacacagtg
     3061 gggttggcac tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac
     3121 ccgaattgcc ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc
     3181 tgccttctgg tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt
     3241 cccattgccg ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg
     3301 cccttctttg acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac
     3361 gttaattaat tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg
     3421 gggctgatac agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca
     3481 cggaggtccc catgtgctgc agtgtcaccg cctccgagga gacagtattg tccctgcggt
     3541 gtccctgcag ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag
     3601 ccccgatgca acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac
     3661 acagggctgg gtcccgctcc cctaatatct acagtgcttc ttgcatggcc ccttaatcaa
     3721 tgcagttaat cagcatgcgc tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct
     3781 gctcaccaac accgcgcacc gccccggccc agcctgcagc acgcgctgca aacaggaaag
     3841 aaacaaaata ttgcccaaat gtaggcaaag gcattcggct gccttgacct ccgccgggcc
     3901 gggccctgcc tgactcagct ccttactcag cgctcgcttc ctccctccgg ctgccaccgc
     3961 cgcagcgcac accctgacaa agagtggccc ttaacgggct ctgaggtgca cccagcagtg
     4021 cactcagcag tccaagggcc ggcctggagg tttgcaccgc tacgtgctga cattagcatt
     4081 gaacttggcc ctgggtagtg ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc
     4141 gttgcacccg gtgccccttc ccctcccttg catcccagca ggctgcaccc cagcaccagg
     4201 cccgtgcatg catgctcctg gtgttattgc agcctggtgc atgcatgcgt cttgatggtg
     4261 cagcgctgtg catgcatcct ccttggtgtg tagcagtcta gtgcatgcat acccctcggt
     4321 gttattgctg ctctgtgcac gcacgctcat tgtatcactt catcccagtg catgcactca
     4381 cactggagcg attgctgctc ggtgcacgca cactcattgt atcactgcag ctcagtggct
     4441 gcacgcacac cggtgttatt gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct
     4501 cagtgcatgc acgctcattg cccatcgcta tccctgcctc tcctgctggc gctccccggg
     4561 aggtgacttc aaggggaccg caggaccacc tcgggggtgg ggggagggct gcacacgcgg
     4621 accccgctcc ccctccccaa caaagcactg tggaatcaaa aaggggggag gggggatgga
     4681 ggggcgcgtc acacccccgc cccacaccct cacctcgagg tgagccccac gttctgcttc
     4741 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     4801 ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     4861 cgrggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     4921 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     4981 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     5041 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     5101 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
     5161 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
     5221 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
     5281 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
     5341 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
     5401 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac
     5461 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     5521 cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg ggkggcggca ggkgggggtg
     5581 ccgggcgggg cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc
     5641 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
     5701 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
     5761 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
     5821 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct
     5881 cggggctgtc cgcaggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     5941 ttctggcgtg tgaccggcgg ggtttatatc ttcccttctc tgttcctccg cagcccccaa
     6001 gctagcatgg cttctagaat cctctggcat gagatgtggc atgaaggcct ggaagaggca
     6061 tctcgtttgt actttgggga aaggaacgtg aaaggcatgt ttgaggtgct ggagcccttg
     6121 catgctatga tggaacgggg cccccagact ctgaaggaaa catcctttaa tcaggcctat
     6181 ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt acatgaaatc agggaatgtc
     6241 aaggacctcc tccaagcctg ggacctctat tatcatgtgt tccgacgaat ctcaaaggga
     6301 agttcaggag gttctagtat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     6361 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     6421 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     6481 cccgtgccct ggcccaccct cgtgaccacc ctgagctggg gcgtgcagtg cttcgcccgc
     6541 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     6601 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     6661 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     6721 ggcaacatcc tggggcacaa gctggagtac aacgccatcc acggcaacgt ctatatcacc
     6781 gccgacaagc agaagaacgg catcaaggcc aacttcggcc tcaactgcaa catcgaggac
     6841 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     6901 ctgctgcccg acaaccacta cctgagcacc cagtccgcac tgagcaaaga ccccaacgag
     6961 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggtctgg tggatcaagt
     7021 ggtggaggta aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtagaagctt
     7081 cgaattctgc agtcg
//
