LOCUS       sequence-107629-        7062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rab5a with N-terminal FRB and triple myc tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Bentley M, Decker H, Luisi J, Banker G
  TITLE     A novel assay reveals preferential binding between Rabs, kinesins, 
            and specific endosomal subpopulations.
  JOURNAL   J Cell Biol. 2015 Feb 2;208(3):273-281. Epub 2015 Jan 26.
  PUBMED    25624392
REFERENCE   2  (bases 1 to 7062)
  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: "J Cell 
            Biol."; date: "2015-02-2"; volume: "208(3)"; pages: "273-281. Epub 
            2015 Jan 26"
COMMENT     Sequence Label: pBa-FRB-3myc-Rab5a
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        418..693
                     /label=chicken beta-actin promoter
     primer_bind     1626..1648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1728..2006
                     /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    2899..3033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2936..2955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2990..3009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3065..3081)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3065..3081)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3078..3100)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3089..3105
                     /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(3113..3119,3120..3137,3138..3143))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3158..3179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3296..3313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3467..4055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3547..3566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4226..5017,5018..5086))
                     /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     4849..4868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5087..5191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5259..5277
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5315..5337)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5437..5456
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5650..5672
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5664..5681
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5665..5681
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct cattgtatca
       61 cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac gcacactcat
      121 tgtatcacgt cagctcagtg gctgcacgca caccggtgtt attgctgctc ggtgcgtgca
      181 tgcacatcag tgtcgctgca gctcagtgca tgcacgctca ttgcccatcg ctatccctgc
      241 ctctcctgct ggcgctcccc gggaggtgac ttcaagggga ccgcaggacc acctcggggg
      301 tggggggagg gctgcacacg cggaccccgc tccccctccc caacaaagca ctgtggaatc
      361 aaaaaggggg gaggggggat ggaggggcgc gtcacacccc cgccccacac cctcacctcg
      421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
      541 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      601 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      661 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
      721 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      781 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
      841 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
      901 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
      961 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1021 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     1081 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1141 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1201 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1261 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1321 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1381 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     1441 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     1501 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1561 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     1621 ggcggggttc ggcttctggc gtgtgaccgg cggggtttat atcttccctt ctctgttcct
     1681 ccgcagcccc caagcttggc gcgcctaggc ctaccatggc ttctagaatc ctctggcatg
     1741 agatgtggca tgaaggcctg gaagaggcat ctcgtttgta ctttggggaa aggaacgtga
     1801 aaggcatgtt tgaggtgctg gagcccttgc atgctatgat ggaacggggc ccccagactc
     1861 tgaaggaaac atcctttaat caggcctatg gtcgagattt aatggaggcc caagagtggt
     1921 gcaggaagta catgaaatca gggaatgtca aggacctcct ccaagcctgg gacctctatt
     1981 atcatgtgtt ccgacgaatc tcaaagacta gttatccgta cggaatgcag aagctgatct
     2041 cagaggagga cctgcaaaaa cttatttctg aagaggatct gcaaaagctt atttctgaag
     2101 aagatctgta caagcggacc ggaagtggaa tggctaatcg aggagcaaca agacccaacg
     2161 ggccaaatac tggaaataaa atatgccagt tcaaactggt ccttctagga gagtctgctg
     2221 ttggcaaatc aagcctggtt cttcgctttg tgaaaggcca atttcatgaa tttcaagaga
     2281 gtaccattgg ggctgccttt ctaacccaaa ctgtgtgtct tgatgacaca acagtaaaat
     2341 ttgaaatatg ggatacagct ggtcaagaac ggtatcatag cttagcacca atgtactacc
     2401 gaggagcaca agcagccata gttgtgtatg atatcacaaa tgaggaatcc tttgcgagag
     2461 caaaaaactg ggttaaagaa cttcaaaggc aagcaagtcc taatattgtg atagctttgt
     2521 caggaaacaa agctgactta gcaaataaaa gagctgttga cttccaggaa gcacagtcct
     2581 atgcagatga caacagctta ttatttatgg agacatcagc taagacatca atgaatgtaa
     2641 atgaaatatt tatggcaata gctaaaaagc tgccaaagaa tgaaccacag aatcctggtg
     2701 caaactcagc cagaggacga ggagtagacc ttactgagcc tgcacagcca gccagaagcc
     2761 agtgttgtag taactgattc tagataagta atgatcataa tcagccatat cacatctgta
     2821 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2881 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2941 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3001 aaactcatca atgtatctta tcatgtctgg atccccgggt accgagctcg aattcgtaat
     3061 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3121 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3181 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3241 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3301 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3361 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3901 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5221 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5281 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5341 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5401 cgggtgttgg cgggtgtcgg ggttggctta actatgcggc atcagagcag attgtactga
     5461 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     5521 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5581 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     5641 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagttg ggatctttgc
     5701 attggcccac ggctctcagg atggggatgc tccccttcag cacccggttc cccttggaaa
     5761 ctgatggtcc tggctctgtg gcatggcagt ggcactgtga ggagccccta ccagcagcac
     5821 acagtgggtt tggcactgcc acgctccgga tgccgcgctc tgatccaacc ccataatcaa
     5881 gggaacccga attgccccat cattgccccc accaccccca tcctgccggg ccctcacacc
     5941 ccacgctgcc ttgtggtgac attccccagc ccaaacccac ggcttcatgg ctaccgcggg
     6001 gcatttccca ttgccgcccc attatcagct ctgcacacct cccgctgtac ccatgcctcg
     6061 tggctgccct tctttgacgt ataatcttct aattaatacc cggccttgtc aaagtggagc
     6121 acaaacgtta attaattccc cagcaggcag gtaattaaca gtgtgactcc ctttttgctg
     6181 cgagtggggc tgatacagag agatgtggca ctatggagcc cacggggtcc tggcactggg
     6241 tgcccacgga ggtccccatg tgctgcagtg tcaccgcctc cgaggtgaca gtattgtccc
     6301 tgcggtgtcc ctgcagctca gctctgtcca cagggccacc tccagtttgg aggggacaca
     6361 atgcagcccc gatgcaaccc atcctcgcag catcccaggg acaaagaccc cactgcaaga
     6421 ccgcacacag ggctgggtcc cgctccccta atatctacag tgcttttgca tggcccctta
     6481 atcaatgcag ttaatcagca tgcgctcatg caccgctctg gagctgcaaa gcccctcgca
     6541 gcgctgctca ccaacaccgc gcaccgcccc ggcccagcct gcagcacgcg ctgcaaacag
     6601 gaaagaaaca aaatattgcc caaatgtagg caaaggcatt cggctgcctt gacctccgcc
     6661 gggccgggcc ctgcctgact cagctcctta ctcagcgctc gcttcctccc tccggctgcc
     6721 accgccgcag cgcacaccct gacaaagagt ggcccttaac gggctctgag gtgcacccag
     6781 cagtgcactc agcagtccaa gggccggcct ggaggtttgc accgctacgt gctgacatta
     6841 gcattgaact tggccctggg tagtgctgca ggccgggcgg ggtgggtgta gagagtgcag
     6901 cgcgcgttgc acccggtgcc ccttcccctc ccttgcatcc cagcaggctg caccccagca
     6961 ccaggcccgt gcatgcatgc tcctggtgtt attgcagcct ggtgcatgca tgcgtcttag
     7021 tggtgcagcg ctgtgcatgc atcctccttg gtgtgtagca gc
//
