LOCUS       sequence-383743-        7493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapalog-inducible coupling to plasma membrane (CAAX) via FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  AUTHORS   Burute M, Jansen KI, Mihajlovic M, Vermonden T, Kapitein LC
  TITLE     Local changes in microtubule network mobility instruct neuronal 
            polarization and axon specification.
  JOURNAL   Sci Adv. 2022 Nov 4;8(44):eabo2343. doi: 10.1126/sciadv.abo2343. 
            Epub 2022 Nov 4.
  PUBMED    36332030
REFERENCE   2  (bases 1 to 7493)
  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):eabo2343. doi"; pages: " 
            10.1126/sciadv.abo2343. Epub 2022 Nov 4"
COMMENT     Sequence Label: pB80-2xFKBP-L-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..7493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     941..963
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1026..1035
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1032..1034,1035..1355)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             1362..1682
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1362..1682
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1788..2492
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1930..1948)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2652..2786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2689..2708)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2743..2762
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2800..2816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2800..2816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2813..2835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2824..2840
                     /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(2848..2854,2855..2872,2873..2878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2893..2914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3031..3048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3202..3790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3282..3301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3961..4752,4753..4821))
                     /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     4584..4603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4822..4926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4994..5012
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5050..5072)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5172..5191
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5385..5407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5399..5416
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5400..5416
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7217..7491
                     /label=chicken beta-actin promoter
ORIGIN
        1 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
       61 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      121 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      181 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      241 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      301 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
      361 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
      421 gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac
      481 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tgcggggcgt
      541 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg kgggggtgcc gggcggggcg
      601 gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
      661 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      721 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      781 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      841 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccatc tccagcctcg gggctgtccg
      901 cagggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      961 accggcgggg tttatatctt cccttctctg ttcctccgca gcccccaagc taggcaattg
     1021 ggcgcgccac catgggagtg caggtggaaa ccatctcccc aggagacggg cgcaccttcc
     1081 ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat ggaaagaaat
     1141 ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag caggaggtga
     1201 tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc aaactgacta
     1261 tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca ccacatgcca
     1321 ctctcgtctt cgatgtggag cttctaaaac tggaaactag aggcgtgcag gtcgagacca
     1381 tcagccccgg cgacggccgc acctttccca agagaggcca gacttgcgtg gtccactaca
     1441 ccggcatgct ggaggacggc aagaagttcg acagcagccg cgaccgcaac aagcccttca
     1501 agttcatgct gggcaaacag gaagtgatcc gcggctggga ggaaggcgtg gctcagatga
     1561 gcgtggggca gcgggccaag ctgaccatca gccccgacta tgcctacggc gccaccggcc
     1621 accccggcat catccccccc cacgccaccc tggtgttcga cgtggagctg ctgaagctgg
     1681 agactagttc tagaggtggc gggggcagtg ggggaggggg aagtggaggc ggcggttctg
     1741 gcgggggagg ctccggtggc ggggggagcg ggggtgggag taagcttgtg agcaagggcg
     1801 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     1861 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1921 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1981 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2041 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2101 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2161 aggtgaagct gcgcggcacc aacttcccct ccgacggtcc cgtaatgcag aagaagacca
     2221 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2281 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2341 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2401 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2461 actccaccgg cggcatggac gagctgtaca aggctagcta cgtatgtgta attatgtgaa
     2521 cgcgtaccgg tcctagataa gtaatgatca taatcagcca tatcacatct gtagaggttt
     2581 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2641 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2701 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2761 tcaatgtatc ttatcatgtc tggatcaatt cgtaatcatg tcatagctgt ttcctgtgtg
     2821 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2881 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2941 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3001 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3061 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3121 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3181 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3241 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3301 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3361 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3421 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3481 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3541 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3601 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3661 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3721 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3781 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3841 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3901 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3961 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4021 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4081 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4141 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4201 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4261 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4321 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4381 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4441 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4501 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4561 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4621 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4681 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4741 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4801 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4861 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4921 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4981 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5041 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5101 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5161 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5221 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5281 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5341 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     5401 taaaacgacg gccagtgcca agttgggatc tttgcattgg cccacggctc tcaggatggg
     5461 gatgctcccc ttcagcaccc ggttcccctt ggaaactgat ggtcctggct ctgtggcatg
     5521 gcagtggcac tgtgaggagc cccctaccag cagcacacag tggggttggc actgccacgc
     5581 tccggatgcc gcgctctgat ccaaccccat aatcaaggga acccgaattg ccccatcatt
     5641 gcccccacca cccccatcct gccgggccct cacaccccac gctgccttct ggtgacattc
     5701 cccagcccaa acccacggct tcatggctac cgcggggcat ttcccattgc cgccccatta
     5761 tcagctctgc acacctcccg ctgtacccat gcctcgtggc tgcccttctt tgacgtataa
     5821 tcttctaatt aatacccggc cttgtcaaag tggagcacaa acgttaatta attccccagc
     5881 aggcaggtaa ttaacagtgt gactcccttt ttgctgcgag tggggctgat acagagagat
     5941 gtggcactat ggagcccacg gggtcctggc actgggtgcc cacggaggtc cccatgtgct
     6001 gcagtgtcac cgcctccgag gagacagtat tgtccctgcg gtgtccctgc agctcagctc
     6061 tgtccacagg gccacctcca gtttggaggg gacacaatgc agccccgatg caacccatcc
     6121 tcgcagcatc ccagggacaa agaccccact gcaagaccgc acacagggct gggtcccgct
     6181 cccctaatat ctacagtgct tcttgcatgg ccccttaatc aatgcagtta atcagcatgc
     6241 gctcatgcac cgctctggag ctgcaaagcc cctcgcagcg ctgctcacca acaccgcgca
     6301 ccgccccggc ccagcctgca gcacgcgctg caaacaggaa agaaacaaaa tattgcccaa
     6361 atgtaggcaa aggcattcgg ctgccttgac ctccgccggg ccgggccctg cctgactcag
     6421 ctccttactc agcgctcgct tcctccctcc ggctgccacc gccgcagcgc acaccctgac
     6481 aaagagtggc ccttaacggg ctctgaggtg cacccagcag tgcactcagc agtccaaggg
     6541 ccggcctgga ggtttgcacc gctacgtgct gacattagca ttgaacttgg ccctgggtag
     6601 tgctgcaggc cgggcggggt gggtgtagag agtgcagcgc gcgttgcacc cggtgcccct
     6661 tcccctccct tgcatcccag caggctgcac cccagcacca ggcccgtgca tgcatgctcc
     6721 tggtgttatt gcagcctggt gcatgcatgc gtcttgatgg tgcagcgctg tgcatgcatc
     6781 ctccttggtg tgtagcagtc tagtgcatgc atacccctcg gtgttattgc tgctctgtgc
     6841 acgcacgctc attgtatcac ttcatcccag tgcatgcact cacactggag cgattgctgc
     6901 tcggtgcacg cacactcatt gtatcactgc agctcagtgg ctgcacgcac accggtgtta
     6961 ttgctgctcg gtgcgtgcat gcacatcagt gtcgctgcag ctcagtgcat gcacgctcat
     7021 tgcccatcgc tatccctgcc tctcctgctg gcgctccccg ggaggtgact tcaaggggac
     7081 cgcaggacca cctcgggggt ggggggaggg ctgcacacgc ggaccccgct ccccctcccc
     7141 aacaaagcac tgtggaatca aaaagggggg aggggggatg gaggggcgcg tcacaccccc
     7201 gccccacacc ctcacctcga ggtgagcccc acgttctgct tcactctccc catctccccc
     7261 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     7321 gcgggggggg gggggcgcgc gccaggsggg gsgggggggg gsggggggsg gggcggggcg
     7381 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     7441 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc ggg
//
