LOCUS       sequence-345338-        8551 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 8551)
  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 8551)
  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..8551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1059..1081
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1144..1153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1148..1157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1150..1233,1234..2400)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     CDS             2500..3204
                     /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(2642..2660)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             3304..3741
                     /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    3871..4005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3908..3927)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3962..3981
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4019..4035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4019..4035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4032..4054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4043..4059
                     /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(4067..4073,4074..4091,4092..4097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4112..4133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4250..4267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4421..5009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4501..4520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5180..5971,5972..6040))
                     /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     5803..5822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6041..6145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6213..6231
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6269..6291)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6391..6410
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6604..6626
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6618..6635
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6619..6635
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc
      301 ttaaagggct ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg
      361 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      421 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
      481 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
      541 gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
      601 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
      661 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
      721 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
      781 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
      841 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
      901 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
      961 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgtccgca
     1021 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1081 cggcggggtt tatatcttcc cttctctgtt cctccgcagc ccccaagcta ggcaattggg
     1141 cgcgccacca tggcggcccc cggcagcgcc cggcgacccc tgctgctgct actgctgttg
     1201 ctgctgctcg gcctcatgca ttgtgcgtca gcagcaatgt ttatggtgaa aaatggcaac
     1261 gggaccgcgt gcataatggc caacttctct gctgccttct cagtgaacta cgacaccaag
     1321 agtggcccta agaacatgac ctttgacctg ccatcagatg ccacagtggt gctcaaccgc
     1381 agctcctgtg gaaaagagaa cacttctgac cccagtctcg tgattgcttt tggaagagga
     1441 catacactca ctctcaattt cacgagaaat gcaacacgtt acagcgtcca gctcatgagt
     1501 tttgtttata acttgtcaga cacacacctt ttccccaatg cgagctccaa agaaatcaag
     1561 actgtggaat ctataactga catcagggca gatatagata aaaaatacag atgtgttagt
     1621 ggcacccagg tccacatgaa caacgtgacc gtaacgctcc atgatgccac catccaggcg
     1681 tacctttcca acagcagctt cagcagggga gagacacgct gtgaacaaga caggccttcc
     1741 ccaaccacag cgccccctgc gccacccagc ccctcgccct cacccgtgcc caagagcccc
     1801 tctgtggaca agtacaacgt gagcggcacc aacgggacct gcctgctggc cagcatgggg
     1861 ctgcagctga acctcaccta tgagaggaag gacaacacga cggtgacaag gcttctcaac
     1921 atcaacccca acaagacctc ggccagcggg agctgcggcg cccacctggt gactctggag
     1981 ctgcacagcg agggcaccac cgtcctgctc ttccagttcg ggatgaatgc aagttctagc
     2041 cggtttttcc tacaaggaat ccagttgaat acaattcttc ctgacgccag agaccccgcc
     2101 tttaaagctg ccaacggctc cctgcgagcg ctgcaggcca cagtcggcaa ttcctacaag
     2161 tgcaacgcgg aggagcacgt ccgtgtcacg aaggcgtttt cagtcaatat attcaaagtg
     2221 tgggtccagg ctttcaaggt ggaaggtggc cagtttggct ctgtggagga gtgtctgctg
     2281 gacgagaaca gcatgctgat ccccatcgct gtgggtggtg ccctggcggg gctggtcctc
     2341 atcgtcctca tcgcctacct cgtcggcagg aagaggagtc acgcaggcta ccagactatc
     2401 tctagaggtg gcgggggcag tgggggaggg ggaagtggag gcggcggttc tggcggggga
     2461 ggctccggtg gcggggggag cgggggtggg agtaagcttg tgagcaaggg cgaggaggat
     2521 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     2581 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2641 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     2701 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     2761 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     2821 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     2881 ctgcgcggca ccaacttccc ctccgacggt cccgtaatgc agaagaagac catgggctgg
     2941 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3001 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3061 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3121 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3181 ggcggcatgg acgagctgta caaggctagc ggaggcggtg gctcaggcgg aggagggtca
     3241 ggagggggag gcagtggcgg gggcggctct ggaggagggg gttctggtgg tggtagctac
     3301 gtactggcaa ccacactgga acggatcgag aaaaatttcg tgattactga tccgagactg
     3361 cctgacaacc caatcatttt tgcgagcgat tccttcctgc agctgacaga atattctcgg
     3421 gaagagatcc tggggcgcaa ttgccgtttt ctgcagggac ccgagacaga ccgtgccact
     3481 gttcggaaaa tcagagatgc tattgacaac cagactgaag tgaccgttca gctgatcaat
     3541 tataccaaga gcggcaagaa gttctggaac gtgttccacc tgcagccgat gcgcgattat
     3601 aagggcgacg tccagtactt cattggcgtg cagctggatg gcaccgaacg tcttcatggc
     3661 gccgctgagc gtgaggcggt ctgcctgatc aaaaagacag cctttcagat tgctgaggca
     3721 gcgaacgacg aaaattactt ttgaaccggt cctagataag taatgatcat aatcagccat
     3781 atcacatctg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3841 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3901 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3961 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaattc gtaatcatgt
     4021 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4081 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4141 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4201 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4261 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4321 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5161 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5341 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5401 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5461 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5581 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5701 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5821 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5881 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6061 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6121 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6181 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     6241 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     6301 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     6361 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     6421 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     6481 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6541 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6601 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gttgggatct ttgcattggc
     6661 ccacggctct caggatgggg atgctcccct tcagcacccg gttccccttg gaaactgatg
     6721 gtcctggctc tgtggcatgg cagtggcact gtgaggagcc ccctaccagc agcacacagt
     6781 ggggttggca ctgccacgct ccggatgccg cgctctgatc caaccccata atcaagggaa
     6841 cccgaattgc cccatcattg cccccaccac ccccatcctg ccgggccctc acaccccacg
     6901 ctgccttctg gtgacattcc ccagcccaaa cccacggctt catggctacc gcggggcatt
     6961 tcccattgcc gccccattat cagctctgca cacctcccgc tgtacccatg cctcgtggct
     7021 gcccttcttt gacgtataat cttctaatta atacccggcc ttgtcaaagt ggagcacaaa
     7081 cgttaattaa ttccccagca ggcaggtaat taacagtgtg actccctttt tgctgcgagt
     7141 ggggctgata cagagagatg tggcactatg gagcccacgg ggtcctggca ctgggtgccc
     7201 acggaggtcc ccatgtgctg cagtgtcacc gcctccgagg agacagtatt gtccctgcgg
     7261 tgtccctgca gctcagctct gtccacaggg ccacctccag tttggagggg acacaatgca
     7321 gccccgatgc aacccatcct cgcagcatcc cagggacaaa gaccccactg caagaccgca
     7381 cacagggctg ggtcccgctc ccctaatatc tacagtgctt cttgcatggc cccttaatca
     7441 atgcagttaa tcagcatgcg ctcatgcacc gctctggagc tgcaaagccc ctcgcagcgc
     7501 tgctcaccaa caccgcgcac cgccccggcc cagcctgcag cacgcgctgc aaacaggaaa
     7561 gaaacaaaat attgcccaaa tgtaggcaaa ggcattcggc tgccttgacc tccgccgggc
     7621 cgggccctgc ctgactcagc tccttactca gcgctcgctt cctccctccg gctgccaccg
     7681 ccgcagcgca caccctgaca aagagtggcc cttaacgggc tctgaggtgc acccagcagt
     7741 gcactcagca gtccaagggc cggcctggag gtttgcaccg ctacgtgctg acattagcat
     7801 tgaacttggc cctgggtagt gctgcaggcc gggcggggtg ggtgtagaga gtgcagcgcg
     7861 cgttgcaccc ggtgcccctt cccctccctt gcatcccagc aggctgcacc ccagcaccag
     7921 gcccgtgcat gcatgctcct ggtgttattg cagcctggtg catgcatgcg tcttgatggt
     7981 gcagcgctgt gcatgcatcc tccttggtgt gtagcagtct agtgcatgca tacccctcgg
     8041 tgttattgct gctctgtgca cgcacgctca ttgtatcact tcatcccagt gcatgcactc
     8101 acactggagc gattgctgct cggtgcacgc acactcattg tatcactgca gctcagtggc
     8161 tgcacgcaca ccggtgttat tgctgctcgg tgcgtgcatg cacatcagtg tcgctgcagc
     8221 tcagtgcatg cacgctcatt gcccatcgct atccctgcct ctcctgctgg cgctccccgg
     8281 gaggtgactt caaggggacc gcaggaccac ctcgggggtg gggggagggc tgcacacgcg
     8341 gaccccgctc cccctcccca acaaagcact gtggaatcaa aaagggggga ggggggatgg
     8401 aggggcgcgt cacacccccg ccccacaccc tcacctcgag gtgagcccca cgttctgctt
     8461 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     8521 attttgtgca gcgatggggg cggggggggg g
//
