LOCUS       sequence-329886-        6813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of rat LAMP1 fused with mEmerald to perform live tracking
            of lysosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6813)
  AUTHORS   Courchet J, Lewis TL Jr, Lee S, Courchet V, Liou DY, Aizawa S, 
            Polleux F
  TITLE     Terminal axon branching is regulated by the LKB1-NUAK1 kinase 
            pathway via presynaptic mitochondrial capture.
  JOURNAL   Cell. 2013 Jun 20;153(7):1510-25. doi: 10.1016/j.cell.2013.05.021.
  PUBMED    23791179
REFERENCE   2  (bases 1 to 6813)
  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"; doi: 
            "10.1016/j.cell.2013.05"; journalName: "Cell"; date: "2013-06-20- 
            20"; volume: "153"; issue: "7"; pages: "1510-25"
COMMENT     Sequence Label: pCAG mEmerald-LAMP1
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          127..1143
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1067..1089
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1151..1170
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1205..1214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1209..1218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1211..1273,1274..2431)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      2486..2495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2492..2494,2495..2497,2498..3211)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2537..2558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2798..2817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3145..3166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3302..3321)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3367..3422
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3421..3440)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3813..3829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3813..3829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3826..3848)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3837..3853
                     /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(3861..3867,3868..3885,3886..3891))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3906..3927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3986..4181
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4032..4167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4187..4321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4224..4243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4278..4297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4389..4406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4560..5148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4640..4659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5319..6110,6111..6179))
                     /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     5942..5961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6180..6284)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6315..6694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      181 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      241 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      301 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      361 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      421 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
      481 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
      541 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
      601 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
      661 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
      721 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
      781 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
      841 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
      901 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
      961 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1021 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1081 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1141 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgctc
     1201 gagcgccacc atggcggccc cgggcgcccg gcgtccgctg ctcctgttgc tgctggcagg
     1261 ccttgcacac agcgccccag cactgttcga ggtgaaagac aacaacggca cagcgtgtat
     1321 aatggccagc ttctctgcct cctttctgac cacctatgag gctggacatg tttctaaggt
     1381 ctcgaatatg accctgccag cctctgcaga agtcctgaag aatagcagct cttgtggtga
     1441 aaagaatgct tctgagccca ccctcgcaat cacctttgga gaaggatatt tactgaaact
     1501 caccttcaca aaaaacacaa cacgttacag tgtccagcac atgtatttca catataacct
     1561 gtcagacaca caattctttc ccaatgccag ctccaaaggg cccgacactg tggattccac
     1621 aactgacatc aaggcagaca tcaacaaaac ataccgatgt gtcagcgaca tcagggtcta
     1681 catgaagaat gtgaccattg tgctctggga cgctactatc caggcctacc tgccgagtag
     1741 caacttcagc aaggaagaga cacgctgccc acaggatcaa ccttccccaa ctactgggcc
     1801 acccagcccc tcaccaccac ttgtgcccac aaaccccagt gtgtccaagt acaatgtgac
     1861 tggtgacaat ggaacctgcc tgctggcctc tatggcactg caactcaaca tcacctacat
     1921 gaagaaggac aacacgactg tgaccagagc attcaacatc aacccaagtg acaaatatag
     1981 tgggacttgc ggtgcccagt tggtgaccct gaaggtgggg aacaagagca gagtcctgga
     2041 gctgcagttt gggatgaatg ccacttctag cctgtttttc ctgcaaggag ttcagttgaa
     2101 catgactctt cctgatgcca tagagcccac gttcagcacc tccaactatt ccctgaaagc
     2161 tcttcaggcc agtgtcggca actcatacaa gtgcaacagt gaggagcaca tctttgtcag
     2221 caaggcgctc gccctcaatg tcttcagcgt gcaagtccag gctttcaggg tagaaagtga
     2281 caggtttggg tctgtggaag agtgtgtaca ggacggtaac aacatgctga tccccattgc
     2341 tgtgggcggg gccctggcag ggctggtcct catcgtcctc atcgcctacc tcatcggcag
     2401 gaagaggagt cacgcgggct atcagaccat ctcggaattc ggctccaccg gctccaccgg
     2461 ctccaccggc gcggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac
     2521 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2581 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2641 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttgacct acggcgtgca
     2701 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2761 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2821 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2881 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2941 ggtctatatc accgccgaca agcagaagaa cggcatcaag gtgaacttca agacccgcca
     3001 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3061 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     3121 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3181 cactctcggc atggacgagc tgtacaagta agcggccgca attcactcct caggtgcagg
     3241 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     3301 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     3361 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     3421 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     3481 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     3541 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     3601 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     3661 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     3721 tagctgtccc tcttctctta tggagatccc tcgacctgca ccgtcgacca gctggtcgac
     3781 ggtgcaccgt cgaccagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3841 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3901 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3961 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     4021 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4081 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4141 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     4201 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4261 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4321 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6301 gtgccacctg ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     6361 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6421 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6481 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     6541 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6601 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6661 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
     6721 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     6781 attttgtgca gcgatggggg cggggggggg ggg
//
