LOCUS       sequence-347052-        7995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic coupling to early endosomes (RAB5) via iLID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7995)
  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 7995)
  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: "2020-04-6"; volume: "219(4). pii: 133834. doi"; 
            pages: " 10.1083/jcb.201907149"
COMMENT     Sequence Label: pB80-iLID-mCherry-RAB5
FEATURES             Location/Qualifiers
     source          1..7995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     941..963
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1035..1472
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     CDS             1572..2276
                     /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(1714..1732)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3153..3287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3190..3209)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3244..3263
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3301..3317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3301..3317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3314..3336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3325..3341
                     /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(3349..3355,3356..3373,3374..3379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3394..3415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3532..3549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3703..4291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4462..5253,5254..5322))
                     /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     5085..5104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5323..5427)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5495..5513
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5551..5573)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5673..5692
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5886..5908
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5900..5917
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5901..5917
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7718..7993
                     /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 tgggggtgcc gggcggggcg
      601 gggccgcctc gggccgggga 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 catgctggca accacactgg aacggatcga gaaaaatttc gtgattactg
     1081 atccgagact gcctgacaac ccaatcattt ttgcgagcga ttccttcctg cagctgacag
     1141 aatattctcg ggaagagatc ctggggcgca attgccgttt tctgcaggga cccgagacag
     1201 accgtgccac tgttcggaaa atcagagatg ctattgacaa ccagactgaa gtgaccgttc
     1261 agctgatcaa ttataccaag agcggcaaga agttctggaa cgtgttccac ctgcagccga
     1321 tgcgcgatta taagggcgac gtccagtact tcattggcgt gcagctggat ggcaccgaac
     1381 gtcttcatgg cgccgctgag cgtgaggcgg tctgcctgat caaaaagaca gcctttcaga
     1441 ttgctgaggc agcgaacgac gaaaattact tttctagagg tggcgggggc agtgggggag
     1501 ggggaagtgg aggcggcggt tctggcgggg gaggctccgg tggcgggggg agcgggggtg
     1561 ggagtaagct tgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1621 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1681 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1741 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     1801 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     1861 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1921 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     1981 gtcccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     2041 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2101 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2161 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2221 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggcta
     2281 gcggaggcgg tggctcaggc ggaggagggt caggaggggg aggcagtggc gggggcggct
     2341 ctggaggagg gggttctggt ggtggtagct acgtagctag tcgaggcgca acaagaccca
     2401 acgggccaaa tactggaaat aaaatatgcc agttcaaact agtacttctg ggagagtctg
     2461 ctgttggcaa atcaagccta gtgcttcgtt ttgtgaaggg ccaatttcat gaatttcaag
     2521 agagtaccat aggggctgct tttctaaccc aaactgtgtg tcttgatgat acaacagtaa
     2581 agtttgaaat atgggataca gctggtcaag aacgatacca tagcttagca ccaatgtact
     2641 acagaggagc acaagcagcc atagttgtat atgatatcac aaatgaggag tcctttgcca
     2701 gagccaaaaa ctgggttaaa gaacttcaga ggcaagccag tcctaacatt gtaatagctt
     2761 tatcaggaaa caaggctgat cttgcaaata aaagagctgt cgatttccag gaagcacagt
     2821 cctatgcaga tgacaacagt ttattattca tggagacatc agctaaaaca tcgatgaatg
     2881 taaatgaaat attcatggca atagctaaaa agttgccaaa gaacgaacca cagaatccag
     2941 gagcaaattc tgccagagga agaggagtag accttactga acccacgcag ccaaccagga
     3001 atcagtgttg tagtaactaa acgcgtaccg gtcctagata agtaatgatc ataatcagcc
     3061 atatcacatc tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3121 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat tcgtaatcat
     3301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5461 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5521 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5581 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5641 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5701 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5761 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5821 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5881 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagttgggat ctttgcattg
     5941 gcccacggct ctcaggatgg ggatgctccc cttcagcacc cggttcccct tggaaactga
     6001 tggtcctggc tctgtggcat ggcagtggca ctgtgaggag ccccctacca gcagcacaca
     6061 gtggggttgg cactgccacg ctccggatgc cgcgctctga tccaacccca taatcaaggg
     6121 aacccgaatt gccccatcat tgcccccacc acccccatcc tgccgggccc tcacacccca
     6181 cgctgccttc tggtgacatt ccccagccca aacccacggc ttcatggcta ccgcggggca
     6241 tttcccattg ccgccccatt atcagctctg cacacctccc gctgtaccca tgcctcgtgg
     6301 ctgcccttct ttgacgtata atcttctaat taatacccgg ccttgtcaaa gtggagcaca
     6361 aacgttaatt aattccccag caggcaggta attaacagtg tgactccctt tttgctgcga
     6421 gtggggctga tacagagaga tgtggcacta tggagcccac ggggtcctgg cactgggtgc
     6481 ccacggaggt ccccatgtgc tgcagtgtca ccgcctccga ggagacagta ttgtccctgc
     6541 ggtgtccctg cagctcagct ctgtccacag ggccacctcc agtttggagg ggacacaatg
     6601 cagccccgat gcaacccatc ctcgcagcat cccagggaca aagaccccac tgcaagaccg
     6661 cacacagggc tgggtcccgc tcccctaata tctacagtgc ttcttgcatg gccccttaat
     6721 caatgcagtt aatcagcatg cgctcatgca ccgctctgga gctgcaaagc ccctcgcagc
     6781 gctgctcacc aacaccgcgc accgccccgg cccagcctgc agcacgcgct gcaaacagga
     6841 aagaaacaaa atattgccca aatgtaggca aaggcattcg gctgccttga cctccgccgg
     6901 gccgggccct gcctgactca gctccttact cagcgctcgc ttcctccctc cggctgccac
     6961 cgccgcagcg cacaccctga caaagagtgg cccttaacgg gctctgaggt gcacccagca
     7021 gtgcactcag cagtccaagg gccggcctgg aggtttgcac cgctacgtgc tgacattagc
     7081 attgaacttg gccctgggta gtgctgcagg ccgggcgggg tgggtgtaga gagtgcagcg
     7141 cgcgttgcac ccggtgcccc ttcccctccc ttgcatccca gcaggctgca ccccagcacc
     7201 aggcccgtgc atgcatgctc ctggtgttat tgcagcctgg tgcatgcatg cgtcttgatg
     7261 gtgcagcgct gtgcatgcat cctccttggt gtgtagcagt ctagtgcatg catacccctc
     7321 ggtgttattg ctgctctgtg cacgcacgct cattgtatca cttcatccca gtgcatgcac
     7381 tcacactgga gcgattgctg ctcggtgcac gcacactcat tgtatcactg cagctcagtg
     7441 gctgcacgca caccggtgtt attgctgctc ggtgcgtgca tgcacatcag tgtcgctgca
     7501 gctcagtgca tgcacgctca ttgcccatcg ctatccctgc ctctcctgct ggcgctcccc
     7561 gggaggtgac ttcaagggga ccgcaggacc acctcggggg tggggggagg gctgcacacg
     7621 cggaccccgc tccccctccc caacaaagca ctgtggaatc aaaaaggggg gaggggggat
     7681 ggaggggcgc gtcacacccc cgccccacac cctcacctcg aggtgagccc cacgttctgc
     7741 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     7801 ttattttgtg cagcgatggg ggcggggggg gggggggcgc gcgccrggcg gggcggggsg
     7861 gggsgrgggg sggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     7921 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     7981 cgcgcggcgg gcggg
//
