LOCUS       sequence-338794-        7981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic coupling to recycling endosomes (RAB11) via iLID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7981)
  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 7981)
  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-RAB11
FEATURES             Location/Qualifiers
     source          1..7981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     931..953
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1025..1462
                     /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             1562..2266
                     /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(1704..1722)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3140..3274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3177..3196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3231..3250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3289..3305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3289..3305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3302..3324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3313..3329
                     /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(3337..3343,3344..3361,3362..3367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3382..3403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3520..3537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3771..3790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4450..5241,5242..5310))
                     /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     5073..5092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5311..5415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5483..5501
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5539..5561)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5661..5680
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5874..5896
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5888..5905
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5889..5905
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7704..7979
                     /label=chicken beta-actin promoter
ORIGIN
        1 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
       61 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      121 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      181 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      241 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      301 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
      361 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
      421 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
      481 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
      541 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
      601 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
      661 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
      721 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
      781 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
      841 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
      901 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcgggg
      961 tttatatctt cccttctctg ttcctccgca gcccccaagc taggcaattg ggcgcgccac
     1021 catgctggca accacactgg aacggatcga gaaaaatttc gtgattactg atccgagact
     1081 gcctgacaac ccaatcattt ttgcgagcga ttccttcctg cagctgacag aatattctcg
     1141 ggaagagatc ctggggcgca attgccgttt tctgcaggga cccgagacag accgtgccac
     1201 tgttcggaaa atcagagatg ctattgacaa ccagactgaa gtgaccgttc agctgatcaa
     1261 ttataccaag agcggcaaga agttctggaa cgtgttccac ctgcagccga tgcgcgatta
     1321 taagggcgac gtccagtact tcattggcgt gcagctggat ggcaccgaac gtcttcatgg
     1381 cgccgctgag cgtgaggcgg tctgcctgat caaaaagaca gcctttcaga ttgctgaggc
     1441 agcgaacgac gaaaattact tttctagagg tggcgggggc agtgggggag ggggaagtgg
     1501 aggcggcggt tctggcgggg gaggctccgg tggcgggggg agcgggggtg ggagtaagct
     1561 tgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1621 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1681 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1741 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     1801 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     1861 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1921 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gtcccgtaat
     1981 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     2041 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     2101 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     2161 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     2221 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggcta gcggaggcgg
     2281 tggctcaggc ggaggagggt caggaggggg aggcagtggc gggggcggct ctggaggagg
     2341 gggttctggt ggtggtagct acgtaggcac ccgcgacgac gagtacgact atctcttcaa
     2401 agttgtcctt attggagatt ctggtgttgg aaagagtaat ctcttgtctc gatttactcg
     2461 aaatgagttt aatctcgaaa gtaagagcac cattggagta gagtttgcaa caagaagcat
     2521 ccaggttgat gggaaaacaa taaaggcaca gatatgggac acggcagggc aggagcgata
     2581 ccgagctata acatcagcat attatcgtgg agctgtaggt gccttactgg tttatgacat
     2641 tgctaagcat ctcacatatg aaaatgtaga acgatggctg aaagaactga gagatcatgc
     2701 tgatagtaac attgttatca tgcttgtggg caataagagt gatttgcgtc atctcagggc
     2761 agttcctaca gatgaagcaa gagcttttgc agaaaagaat ggtttgtcat tcattgagac
     2821 ttccgctcta gactctacaa atgtagaagc tgcttttcag acaattctga cagagatata
     2881 ccgcattgtt tcccagaaac aaatgtcaga cagacgtgaa aatgacatgt ctccaagcaa
     2941 caatgtggtt cctattcatg ttccaccaac cactgaaaac aagccaaagg tgcagtgctg
     3001 tcagaacatc taaaccggtc ctagataagt aatgatcata atcagccata tcacatctgt
     3061 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3121 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3241 caaactcatc aatgtatctt atcatgtctg gatcaattcg taatcatggt catagctgtt
     3301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacagtagaa ggacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5401 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5461 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5521 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5581 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5641 tcggggttgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5701 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5761 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5821 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5881 acgacgttgt aaaacgacgg ccagtgccaa gttgggatct ttgcattggc ccacggctct
     5941 caggatgggg atgctcccct tcagcacccg gttccccttg gaaactgatg gtcctggctc
     6001 tgtggcatgg cagtggcact gtgaggagcc cctaccagca gcacacagtg ggtttggcac
     6061 tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac ccgaattgcc
     6121 ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc tgccttgtgg
     6181 tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt cccattgccg
     6241 ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg cccttctttg
     6301 acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac gttaattaat
     6361 tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg gggctgatac
     6421 agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca cggaggtccc
     6481 catgtgctgc agtgtcaccg cctccgaggt gacagtattg tccctgcggt gtccctgcag
     6541 ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag ccccgatgca
     6601 acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac acagggctgg
     6661 gtcccgctcc cctaatatct acagtgcttt tgcatggccc cttaatcaat gcagttaatc
     6721 agcatgcgct catgcaccgc tctggagctg caaagcccct cgcagcgctg ctcaccaaca
     6781 ccgcgcaccg ccccggccca gcctgcagca cgcgctgcaa acaggaaaga aacaaaatat
     6841 tgcccaaatg taggcaaagg cattcggctg ccttgacctc cgccgggccg ggccctgcct
     6901 gactcagctc cttactcagc gctcgcttcc tccctccggc tgccaccgcc gcagcgcaca
     6961 ccctgacaaa gagtggccct taacgggctc tgaggtgcac ccagcagtgc actcagcagt
     7021 ccaagggccg gcctggaggt ttgcaccgct acgtgctgac attagcattg aacttggccc
     7081 tgggtagtgc tgcaggccgg gcggggtggg tgtagagagt gcagcgcgcg ttgcacccgg
     7141 tgccccttcc cctcccttgc atcccagcag gctgcacccc agcaccaggc ccgtgcatgc
     7201 atgctcctgg tgttattgca gcctggtgca tgcatgcgtc ttagtggtgc agcgctgtgc
     7261 atgcatcctc cttggtgtgt agcagcttag tgcatgcata cccctcggtg ttattgctgc
     7321 tctgtgcagg cacgctcatt gtatcacttc atcccagtgc atgcactcac actggagcga
     7381 ttgctgctcg gtgcacgcac actcattgta tcacgtcagc tcagtggctg cacgcacacc
     7441 ggtgttattg ctgctcggtg cgtgcatgca catcagtgtc gctgcagctc agtgcatgca
     7501 cgctcattgc ccatcgctat ccctgcctct cctgctggcg ctccccggga ggtgacttca
     7561 aggggaccgc aggaccacct cgggggtggg gggagggctg cacacgcgga ccccgctccc
     7621 cctccccaac aaagcactgt ggaatcaaaa aggggggagg ggggatggag gggcgcgtca
     7681 cacccccgcc ccacaccctc acctcgaggt gagccccacg ttctgcttca ctctccccat
     7741 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     7801 gatgggggcg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     7861 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     7921 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     7981 g
//
