LOCUS       Exported                7875 bp ds-DNA     circular SYN 14-OCT-2021
DEFINITION  Rapalog-inducible coupling to recycling endosomes (RAB11) via FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    pB80-FKBP-mCherry-RAB11
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7875)
  AUTHORS   Nijenhuis W, van Grinsven MMP, Kapitein LC
  TITLE     An optimized toolbox for the optogenetic control of intracellular 
            transport.
  JOURNAL   J Cell Biol. 2020 Apr 6;219(4). pii: 133834. doi: 
            10.1083/jcb.201907149.
  PUBMED    32328628
REFERENCE   2  (bases 1 to 7875)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 14, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7875
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        656..931
                     /label=chicken beta-actin promoter
     primer_bind     1874..1896
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1959..1968
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1965..2288
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             2388..3092
                     /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(2530..2548)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3966..4100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4003..4022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4057..4076
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4114..4130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4114..4130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4127..4149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4138..4154
                     /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(4162..4192)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4207..4228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4345..4362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4516..5104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4596..4615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5275..6135)
                     /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     5898..5917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6136..6240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6308..6326
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6364..6386)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6486..6505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6699..6721
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6713..6730
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6714..6730
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctacgtgctg acattagcat tgaacttggc cctgggtagt gctgcaggcc gggcggggtg
       61 ggtgtagaga gtgcagcgcg cgttgcaccc ggtgcccctt cccctccctt gcatcccagc
      121 aggctgcacc ccagcaccag gcccgtgcat gcatgctcct ggtgttattg cagcctggtg
      181 catgcatgcg tcttgatggt gcagcgctgt gcatgcatcc tccttggtgt gtagcagtct
      241 agtgcatgca tacccctcgg tgttattgct gctctgtgca cgcacgctca ttgtatcact
      301 tcatcccagt gcatgcactc acactggagc gattgctgct cggtgcacgc acactcattg
      361 tatcactgca gctcagtggc tgcacgcaca ccggtgttat tgctgctcgg tgcgtgcatg
      421 cacatcagtg tcgctgcagc tcagtgcatg cacgctcatt gcccatcgct atccctgcct
      481 ctcctgctgg cgctccccgg gaggtgactt caaggggacc gcaggaccac ctcgggggtg
      541 gggggagggc tgcacacgcg gaccccgctc cccctcccca acaaagcact gtggaatcaa
      601 aaagggggga ggggggatgg aggggcgcgt cacacccccg ccccacaccc tcacctcgag
      661 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
      721 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggcgcgc
      781 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
      841 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
      901 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
      961 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     1021 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
     1081 acggctcgtt tcttttctgt ggctgcgtga aagccttaaa gggctccggg agggcccttt
     1141 gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt
     1201 gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc
     1261 tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct
     1321 gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg
     1381 gcgcggcggt cgggctgtaa cccccccctg cacccccctc cccgagttgc tgagcacggc
     1441 ccggcttcgg gtgcggggct ccgtgcgggg cgtggcgcgg ggctcgccgt gccgggcggg
     1501 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
     1561 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     1621 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     1681 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     1741 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     1801 ccccttctcc atctccagcc tcggggctgt ccgcaggggg acggctgcct tcggggggga
     1861 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gggtttatat cttcccttct
     1921 ctgttcctcc gcagccccca agctaggcaa ttgggcgcgc caccatggga gtgcaggtgg
     1981 agactatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
     2041 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
     2101 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
     2161 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
     2221 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
     2281 aactggaatc tagaggtggc gggggcagtg ggggaggggg aagtggaggc ggcggttctg
     2341 gcgggggagg ctccggkggc ggggggrgsg ggggkgggag taagcttgtg agcaagggcg
     2401 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     2461 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2521 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     2581 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2641 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2701 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2761 aggtgaagct gcgcggcacc aacttcccct ccgacggtcc cgtaatgcag aagaagacca
     2821 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2881 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2941 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     3001 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     3061 actccaccgg cggcatggac gagctgtaca aggctagcgg aggcggtggc tcaggcggag
     3121 gagggtcagg agggggaggc agtggcgggg gcggctctgg aggagggggt tctggtggtg
     3181 gtagctacgt aggcacccgc gacgacgagt acgactatct cttcaaagtt gtccttattg
     3241 gagattctgg tgttggaaag agtaatctct tgtctcgatt tactcgaaat gagtttaatc
     3301 tcgaaagtaa gagcaccatt ggagtagagt ttgcaacaag aagcatccag gttgatggga
     3361 aaacaataaa ggcacagata tgggacacgg cagggcagga gcgataccga gctataacat
     3421 cagcatatta tcgtggagct gtaggtgcct tactggttta tgacattgct aagcatctca
     3481 catatgaaaa tgtagaacga tggctgaaag aactgagaga tcatgctgat agtaacattg
     3541 ttatcatgct tgtgggcaat aagagtgatt tgcgtcatct cagggcagtt cctacagatg
     3601 aagcaagagc ttttgcagaa aagaatggtt tgtcattcat tgagacttcc gctctagact
     3661 ctacaaatgt agaagctgct tttcagacaa ttctgacaga gatataccgc attgtttccc
     3721 agaaacaaat gtcagacaga cgtgaaaatg acatgtctcc aagcaacaat gtggttccta
     3781 ttcatgttcc accaaccact gaaaacaagc caaaggtgca gtgctgtcag aacatctaaa
     3841 ccggtcctag ataagtaatg atcataatca gccatatcac atctgtagag gttttacttg
     3901 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3961 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4021 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4081 tatcttatca tgtctggatc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     4141 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4201 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4261 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4321 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4381 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4441 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4921 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6241 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6301 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6361 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6421 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6481 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6541 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6601 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6661 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     6721 gacggccagt gccaagttgg gatctttgca ttggcccacg gctctcagga tggggatgct
     6781 ccccttcagc acccggttcc ccttggaaac tgatggtcct ggctctgtgg catggcagtg
     6841 gcactgtgag gagcccccta ccagcagcac acagtggggt tggcactgcc acgctccgga
     6901 tgccgcgctc tgatccaacc ccataatcaa gggaacccga attgccccat cattgccccc
     6961 accaccccca tcctgccggg ccctcacacc ccacgctgcc ttctggtgac attccccagc
     7021 ccaaacccac ggcttcatgg ctaccgcggg gcatttccca ttgccgcccc attatcagct
     7081 ctgcacacct cccgctgtac ccatgcctcg tggctgccct tctttgacgt ataatcttct
     7141 aattaatacc cggccttgtc aaagtggagc acaaacgtta attaattccc cagcaggcag
     7201 gtaattaaca gtgtgactcc ctttttgctg cgagtggggc tgatacagag agatgtggca
     7261 ctatggagcc cacggggtcc tggcactggg tgcccacgga ggtccccatg tgctgcagtg
     7321 tcaccgcctc cgaggagaca gtattgtccc tgcggtgtcc ctgcagctca gctctgtcca
     7381 cagggccacc tccagtttgg aggggacaca atgcagcccc gatgcaaccc atcctcgcag
     7441 catcccaggg acaaagaccc cactgcaaga ccgcacacag ggctgggtcc cgctccccta
     7501 atatctacag tgcttcttgc atggcccctt aatcaatgca gttaatcagc atgcgctcat
     7561 gcaccgctct ggagctgcaa agcccctcgc agcgctgctc accaacaccg cgcaccgccc
     7621 cggcccagcc tgcagcacgc gctgcaaaca ggaaagaaac aaaatattgc ccaaatgtag
     7681 gcaaaggcat tcggctgcct tgacctccgc cgggccgggc cctgcctgac tcagctcctt
     7741 actcagcgct cgcttcctcc ctccggctgc caccgccgca gcgcacaccc tgacaaagag
     7801 tggcccttaa cgggctctga ggtgcaccca gcagtgcact cagcagtcca agggccggcc
     7861 tggaggtttg caccg
//
