LOCUS       sequence-345346-        7953 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 7953)
  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 7953)
  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..7953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1064..1086
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1158..1595
                     /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             1695..2399
                     /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(1837..1855)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3273..3407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3310..3329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3364..3383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3421..3437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3421..3437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3434..3456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3445..3461
                     /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(3469..3475,3476..3493,3494..3499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3514..3535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3652..3669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3823..4411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3903..3922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4582..5373,5374..5442))
                     /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     5205..5224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5443..5547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5615..5633
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5671..5693)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5793..5812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6006..6028
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6020..6037
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6021..6037
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
       61 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      121 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      181 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      241 tccgggctgt aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga
      301 aagccttaaa gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt
      361 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      421 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
      481 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
      541 tgtgcgtggg ggggtgagca gggggtgtgg gcgcggcggt cgggctgtaa cccccccctg
      601 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtgcgggg
      661 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
      721 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
      781 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
      841 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
      901 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
      961 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc atctccagcc tcggggctgt
     1021 ccgcaggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1081 gtgaccggcg gggtttatat cttcccttct ctgttcctcc gcagccccca agctaggcaa
     1141 ttgggcgcgc caccatgctg gcaaccacac tggaacggat cgagaaaaat ttcgtgatta
     1201 ctgatccgag actgcctgac aacccaatca tttttgcgag cgattccttc ctgcagctga
     1261 cagaatattc tcgggaagag atcctggggc gcaattgccg ttttctgcag ggacccgaga
     1321 cagaccgtgc cactgttcgg aaaatcagag atgctattga caaccagact gaagtgaccg
     1381 ttcagctgat caattatacc aagagcggca agaagttctg gaacgtgttc cacctgcagc
     1441 cgatgcgcga ttataagggc gacgtccagt acttcattgg cgtgcagctg gatggcaccg
     1501 aacgtcttca tggcgccgct gagcgtgagg cggtctgcct gatcaaaaag acagcctttc
     1561 agattgctga ggcagcgaac gacgaaaatt acttttctag aggtggcggg ggcagtgggg
     1621 gagggggaag tggaggcggc ggttctggcg ggggaggctc cggtggcggg gggagcgggg
     1681 gtgggagtaa gcttgtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     1741 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1801 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     1861 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     1921 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     1981 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     2041 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     2101 acggtcccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     2161 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     2221 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     2281 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     2341 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg
     2401 ctagcggagg cggtggctca ggcggaggag ggtcaggagg gggaggcagt ggcgggggcg
     2461 gctctggagg agggggttct ggtggtggta gctacgtagg cacccgcgac gacgagtacg
     2521 actatctctt caaagttgtc cttattggag attctggtgt tggaaagagt aatctcttgt
     2581 ctcgatttac tcgaaatgag tttaatctcg aaagtaagag caccattgga gtagagtttg
     2641 caacaagaag catccaggtt gatgggaaaa caataaaggc acagatatgg gacacggcag
     2701 ggcaggagcg ataccgagct ataacatcag catattatcg tggagctgta ggtgccttac
     2761 tggtttatga cattgctaag catctcacat atgaaaatgt agaacgatgg ctgaaagaac
     2821 tgagagatca tgctgatagt aacattgtta tcatgcttgt gggcaataag agtgatttgc
     2881 gtcatctcag ggcagttcct acagatgaag caagagcttt tgcagaaaag aatggtttgt
     2941 cattcattga gacttccgct ctagactcta caaatgtaga agctgctttt cagacaattc
     3001 tgacagagat ataccgcatt gtttcccaga aacaaatgtc agacagacgt gaaaatgaca
     3061 tgtctccaag caacaatgtg gttcctattc atgttccacc aaccactgaa aacaagccaa
     3121 aggtgcagtg ctgtcagaac atctaaaccg gtcctagata agtaatgatc ataatcagcc
     3181 atatcacatc tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3241 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3301 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3361 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat tcgtaatcat
     3421 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3481 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3541 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3601 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3661 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3721 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3781 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3841 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3901 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3961 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4021 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4081 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4141 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4201 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4261 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4321 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4381 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4441 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4501 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4561 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4621 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4681 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4741 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4801 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4861 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4921 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4981 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5041 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5101 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5161 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5221 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5281 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5341 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5401 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5461 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5521 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5581 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5641 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5701 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5761 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5821 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5881 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5941 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6001 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagttgggat ctttgcattg
     6061 gcccacggct ctcaggatgg ggatgctccc cttcagcacc cggttcccct tggaaactga
     6121 tggtcctggc tctgtggcat ggcagtggca ctgtgaggag ccccctacca gcagcacaca
     6181 gtggggttgg cactgccacg ctccggatgc cgcgctctga tccaacccca taatcaaggg
     6241 aacccgaatt gccccatcat tgcccccacc acccccatcc tgccgggccc tcacacccca
     6301 cgctgccttc tggtgacatt ccccagccca aacccacggc ttcatggcta ccgcggggca
     6361 tttcccattg ccgccccatt atcagctctg cacacctccc gctgtaccca tgcctcgtgg
     6421 ctgcccttct ttgacgtata atcttctaat taatacccgg ccttgtcaaa gtggagcaca
     6481 aacgttaatt aattccccag caggcaggta attaacagtg tgactccctt tttgctgcga
     6541 gtggggctga tacagagaga tgtggcacta tggagcccac ggggtcctgg cactgggtgc
     6601 ccacggaggt ccccatgtgc tgcagtgtca ccgcctccga ggagacagta ttgtccctgc
     6661 ggtgtccctg cagctcagct ctgtccacag ggccacctcc agtttggagg ggacacaatg
     6721 cagccccgat gcaacccatc ctcgcagcat cccagggaca aagaccccac tgcaagaccg
     6781 cacacagggc tgggtcccgc tcccctaata tctacagtgc ttcttgcatg gccccttaat
     6841 caatgcagtt aatcagcatg cgctcatgca ccgctctgga gctgcaaagc ccctcgcagc
     6901 gctgctcacc aacaccgcgc accgccccgg cccagcctgc agcacgcgct gcaaacagga
     6961 aagaaacaaa atattgccca aatgtaggca aaggcattcg gctgccttga cctccgccgg
     7021 gccgggccct gcctgactca gctccttact cagcgctcgc ttcctccctc cggctgccac
     7081 cgccgcagcg cacaccctga caaagagtgg cccttaacgg gctctgaggt gcacccagca
     7141 gtgcactcag cagtccaagg gccggcctgg aggtttgcac cgctacgtgc tgacattagc
     7201 attgaacttg gccctgggta gtgctgcagg ccgggcgggg tgggtgtaga gagtgcagcg
     7261 cgcgttgcac ccggtgcccc ttcccctccc ttgcatccca gcaggctgca ccccagcacc
     7321 aggcccgtgc atgcatgctc ctggtgttat tgcagcctgg tgcatgcatg cgtcttgatg
     7381 gtgcagcgct gtgcatgcat cctccttggt gtgtagcagt ctagtgcatg catacccctc
     7441 ggtgttattg ctgctctgtg cacgcacgct cattgtatca cttcatccca gtgcatgcac
     7501 tcacactgga gcgattgctg ctcggtgcac gcacactcat tgtatcactg cagctcagtg
     7561 gctgcacgca caccggtgtt attgctgctc ggtgcgtgca tgcacatcag tgtcgctgca
     7621 gctcagtgca tgcacgctca ttgcccatcg ctatccctgc ctctcctgct ggcgctcccc
     7681 gggaggtgac ttcaagggga ccgcaggacc acctcggggg tggggggagg gctgcacacg
     7741 cggaccccgc tccccctccc caacaaagca ctgtggaatc aaaaaggggg gaggggggat
     7801 ggaggggcgc gtcacacccc cgccccacac cctcacctcg aggtgagccc cacgttctgc
     7861 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     7921 ttattttgtg cagcgatggg ggcggggggg ggg
//
