LOCUS       sequence-338796-        8584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic coupling to lysosomes (LAMP1) via iLID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8584)
  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 8584)
  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-LAMP1-mCherry-iLID
FEATURES             Location/Qualifiers
     source          1..8584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     931..953
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1016..1025
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1020..1029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1022..1105,1106..2272)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     CDS             2372..3076
                     /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(2514..2532)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2782..2801
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3176..3613
                     /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"
     polyA_signal    3743..3877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3780..3799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3834..3853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3892..3908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3892..3908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3905..3927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3916..3932
                     /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(3940..3946,3947..3964,3965..3970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3985..4006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4123..4140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4294..4882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4374..4393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5053..5844,5845..5913))
                     /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     5676..5695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5914..6018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6086..6104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6142..6164)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6264..6283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6477..6499
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6491..6508
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6492..6508
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8307..8582
                     /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 catggcggcc cccggcagcg cccggcgacc cctgctgctg ctactgctgt tgctgctgct
     1081 cggcctcatg cattgtgcgt cagcagcaat gtttatggtg aaaaatggca acgggaccgc
     1141 gtgcataatg gccaacttct ctgctgcctt ctcagtgaac tacgacacca agagtggccc
     1201 taagaacatg acctttgacc tgccatcaga tgccacagtg gtgctcaacc gcagctcctg
     1261 tggaaaagag aacacttctg accccagtct cgtgattgct tttggaagag gacatacact
     1321 cactctcaat ttcacgagaa atgcaacacg ttacagcgtc cagctcatga gttttgttta
     1381 taacttgtca gacacacacc ttttccccaa tgcgagctcc aaagaaatca agactgtgga
     1441 atctataact gacatcaggg cagatataga taaaaaatac agatgtgtta gtggcaccca
     1501 ggtccacatg aacaacgtga ccgtaacgct ccatgatgcc accatccagg cgtacctttc
     1561 caacagcagc ttcagcaggg gagagacacg ctgtgaacaa gacaggcctt ccccaaccac
     1621 agcgccccct gcgccaccca gcccctcgcc ctcacccgtg cccaagagcc cctctgtgga
     1681 caagtacaac gtgagcggca ccaacgggac ctgcctgctg gccagcatgg ggctgcagct
     1741 gaacctcacc tatgagagga aggacaacac gacggtgaca aggcttctca acatcaaccc
     1801 caacaagacc tcggccagcg ggagctgcgg cgcccacctg gtgactctgg agctgcacag
     1861 cgagggcacc accgtcctgc tcttccagtt cgggatgaat gcaagttcta gccggttttt
     1921 cctacaagga atccagttga atacaattct tcctgacgcc agagaccccg cctttaaagc
     1981 tgccaacggc tccctgcgag cgctgcaggc cacagtcggc aattcctaca agtgcaacgc
     2041 ggaggagcac gtccgtgtca cgaaggcgtt ttcagtcaat atattcaaag tgtgggtcca
     2101 ggctttcaag gtggaaggtg gccagtttgg ctctgtggag gagtgtctgc tggacgagaa
     2161 cagcatgctg atccccatcg ctgtgggtgg tgccctggcg gggctggtcc tcatcgtcct
     2221 catcgcctac ctcgtcggca ggaagaggag tcacgcaggc taccagacta tctctagagg
     2281 tggcgggggc agtgggggag ggggaagtgg aggcggcggt tctggcgggg gaggctccgg
     2341 tggcgggggg agcgggggtg ggagtaagct tgtgagcaag ggcgaggagg ataacatggc
     2401 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     2461 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2521 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     2581 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2641 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2701 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2761 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2821 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2881 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2941 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     3001 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     3061 ggacgagctg tacaaggcta gcggaggcgg tggctcaggc ggaggagggt caggaggggg
     3121 aggcagtggc gggggcggct ctggaggagg gggttctggt ggtggtagct acgtactggc
     3181 aaccacactg gaacggatcg agaaaaattt cgtgattact gatccgagac tgcctgacaa
     3241 cccaatcatt tttgcgagcg attccttcct gcagctgaca gaatattctc gggaagagat
     3301 cctggggcgc aattgccgtt ttctgcaggg acccgagaca gaccgtgcca ctgttcggaa
     3361 aatcagagat gctattgaca accagactga agtgaccgtt cagctgatca attataccaa
     3421 gagcggcaag aagttctgga acgtgttcca cctgcagccg atgcgcgatt ataagggcga
     3481 cgtccagtac ttcattggcg tgcagctgga tggcaccgaa cgtcttcatg gcgccgctga
     3541 gcgtgaggcg gtctgcctga tcaaaaagac agcctttcag attgctgagg cagcgaacga
     3601 cgaaaattac ttttgaaccg gtcctagata agtaatgatc ataatcagcc atatcacatc
     3661 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3721 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3781 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3841 gtccaaactc atcaatgtat cttatcatgt ctggatcaat tcgtaatcat ggtcatagct
     3901 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3961 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4021 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4081 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4201 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacagta gaaggacagt
     4741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6061 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6241 gtgtcggggt tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6361 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6421 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6481 gtcacgacgt tgtaaaacga cggccagtgc caagttggga tctttgcatt ggcccacggc
     6541 tctcaggatg gggatgctcc ccttcagcac ccggttcccc ttggaaactg atggtcctgg
     6601 ctctgtggca tggcagtggc actgtgagga gcccctacca gcagcacaca gtgggtttgg
     6661 cactgccacg ctccggatgc cgcgctctga tccaacccca taatcaaggg aacccgaatt
     6721 gccccatcat tgcccccacc acccccatcc tgccgggccc tcacacccca cgctgccttg
     6781 tggtgacatt ccccagccca aacccacggc ttcatggcta ccgcggggca tttcccattg
     6841 ccgccccatt atcagctctg cacacctccc gctgtaccca tgcctcgtgg ctgcccttct
     6901 ttgacgtata atcttctaat taatacccgg ccttgtcaaa gtggagcaca aacgttaatt
     6961 aattccccag caggcaggta attaacagtg tgactccctt tttgctgcga gtggggctga
     7021 tacagagaga tgtggcacta tggagcccac ggggtcctgg cactgggtgc ccacggaggt
     7081 ccccatgtgc tgcagtgtca ccgcctccga ggtgacagta ttgtccctgc ggtgtccctg
     7141 cagctcagct ctgtccacag ggccacctcc agtttggagg ggacacaatg cagccccgat
     7201 gcaacccatc ctcgcagcat cccagggaca aagaccccac tgcaagaccg cacacagggc
     7261 tgggtcccgc tcccctaata tctacagtgc ttttgcatgg ccccttaatc aatgcagtta
     7321 atcagcatgc gctcatgcac cgctctggag ctgcaaagcc cctcgcagcg ctgctcacca
     7381 acaccgcgca ccgccccggc ccagcctgca gcacgcgctg caaacaggaa agaaacaaaa
     7441 tattgcccaa atgtaggcaa aggcattcgg ctgccttgac ctccgccggg ccgggccctg
     7501 cctgactcag ctccttactc agcgctcgct tcctccctcc ggctgccacc gccgcagcgc
     7561 acaccctgac aaagagtggc ccttaacggg ctctgaggtg cacccagcag tgcactcagc
     7621 agtccaaggg ccggcctgga ggtttgcacc gctacgtgct gacattagca ttgaacttgg
     7681 ccctgggtag tgctgcaggc cgggcggggt gggtgtagag agtgcagcgc gcgttgcacc
     7741 cggtgcccct tcccctccct tgcatcccag caggctgcac cccagcacca ggcccgtgca
     7801 tgcatgctcc tggtgttatt gcagcctggt gcatgcatgc gtcttagtgg tgcagcgctg
     7861 tgcatgcatc ctccttggtg tgtagcagct tagtgcatgc atacccctcg gtgttattgc
     7921 tgctctgtgc aggcacgctc attgtatcac ttcatcccag tgcatgcact cacactggag
     7981 cgattgctgc tcggtgcacg cacactcatt gtatcacgtc agctcagtgg ctgcacgcac
     8041 accggtgtta ttgctgctcg gtgcgtgcat gcacatcagt gtcgctgcag ctcagtgcat
     8101 gcacgctcat tgcccatcgc tatccctgcc tctcctgctg gcgctccccg ggaggtgact
     8161 tcaaggggac cgcaggacca cctcgggggt ggggggaggg ctgcacacgc ggaccccgct
     8221 ccccctcccc aacaaagcac tgtggaatca aaaagggggg aggggggatg gaggggcgcg
     8281 tcacaccccc gccccacacc ctcacctcga ggtgagcccc acgttctgct tcactctccc
     8341 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     8401 agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
     8461 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     8521 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     8581 cggg
//
