LOCUS       sequence-490777-        6197 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Lysosomal marker LAMP1 tagged C-terminally with Halo tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  AUTHORS   Basak B, Holzbaur ELF
  TITLE     Mitochondrial damage triggers the concerted degradation of negative 
            regulators of neuronal autophagy.
  JOURNAL   Nat Commun. 2025 Aug 9;16(1):7367. doi: 10.1038/s41467-025-62379-5.
  PUBMED    40783388
REFERENCE   2  (bases 1 to 6197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-08-9"; volume: "16(1):7367. doi"; pages: " 
            10.1038/s41467-025-62379-5"
COMMENT     Sequence Label: LAMP1-Halo
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     545..564
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(653..1010)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      667..802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        819..1010
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     967..987
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1012..1116)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1143..1598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1230..1249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1440..1461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1605..1726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1616..1635)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1670..1689
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1861..1879
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1951..2835)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             complement(join(2848..4005,4006..4068))
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      4061..4070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4065..4074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4143..4346)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4172..4192)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4347..4650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4825..5413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4905..4924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5742..5789
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5814..5833
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(6021..6197,1..618))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(join(6021..6197,1..618))
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6132..6151)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
       61 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
      121 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
      181 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
      241 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      301 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      361 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
      421 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      481 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      541 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
      601 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
      661 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
      721 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      781 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      841 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      901 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      961 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
     1021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1141 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1561 taacgcgaat tttaacaaaa tattaacgct tacaatttac gccttaagat acattgatga
     1621 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1681 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1741 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1801 cctctacaaa tgtggtatgg ctgattatga tctagagtcg cggccgcccc aaggggttat
     1861 gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc tttgttagca
     1921 gccggatcag cttgcatgcc aattctatta accggaaatc tccagagtag acagccagcg
     1981 cgcgatctcg ctgccgatca ggtccgggtt gtcttcttgc agcagattca gacccgggcc
     2041 gatgtccaca gccttgcagt taggcaggct tttggccagg cgagcggctt cggccggtgg
     2101 gatcagaacg cctggggtgc cccagaacag cagcttcggg acaggggact ggtgcagcca
     2161 gtccatgtat tcttcgacca gcgcgacgat gttcgctggc tcaccggcga ttggcagctc
     2221 gtttgggaag cgccacagtg gctcgcggtc aacaggattc aggaacggct cgcggtaatg
     2281 gtccatctcg acttcagtca gcgggcggac gacacccatc ggcagcgtac cctcgataaa
     2341 aacgttctga tcgatgatca gcttgcggcc gacgtcggtg gtgcggaagg cctggaaggt
     2401 ctcgcgggca aattctggcc attcgtccca ggtcgggata gggcggatga actccataaa
     2461 tgcaatacct ttgacgcgct ctggattgcg cttggcccag tggaaaccca gagcggagcc
     2521 ccagtcgtga atgaccagga cgacctcttc cagacccagg gcttcgatga aggcatccat
     2581 gaagcggacg tggtcgtcga agaaataacc caggtctggt ttgtcggatt tgcccatacc
     2641 gatcaggtct ggagcaatgc agcgatgggt cggtgcaaca tgcgggatga tgttgcgcca
     2701 cacgtaggag gaggtcgggt taccgtgcag gaacagcaca ggggtgccat cgcgcggacc
     2761 aacatcgacg tagtgcatgc gctcgcccag gacttccaca taatgggggt cgaatggaaa
     2821 gccagtaccg atttcggatc gatcccggat ggtctgatag cccgcgtgac tcctcttcct
     2881 gccgatgagg taggcgatga ggacgatgag gaccagccct gccagggccc cgcccacagc
     2941 aatggggatc agcatgttgt taccgtcctg tacacactct tccacagacc caaacctgtc
     3001 actttctacc ctgaaagcct ggacttgcac gctgaagaca ttgagggcga gcgccttgct
     3061 gacaaagatg tgctcctcac tgttgcactt gtatgagttg ccgacactgg cctgaagagc
     3121 tttcagggaa tagttggagg tgctgaacgt gggctctatg gcatcaggaa gagtcatgtt
     3181 caactgaact ccttgcagga aaaacaggct agaagtggca ttcatcccaa actgcagctc
     3241 caggactctg ctcttgttcc ccaccttcag ggtcaccaac tgggcaccgc aagtcccact
     3301 atatttgtca cttgggttga tgttgaatgc tctggtcaca gtcgtgttgt ccttcttcat
     3361 gtaggtgatg ttgagttgca gtgccataga ggccagcagg caggttccat tgtcaccagt
     3421 cacattgtac ttggacacac tggggtttgt gggcacgagt ggtggtgagg ggctgggtgg
     3481 cccagtagtt ggggaaggtt gatcctgtgg gcagcgtgtc tcttccttgc tgaagttgct
     3541 actcggcagg taggcctgga tagtagcgtc ccagagcaca atggtcacat tcttcatgta
     3601 gaccctgatg tcgctgacac atcggtatgt tttgttgatg tctgccttga tgtcagttgt
     3661 ggaatccaca gtgtcgggcc ctttggagct ggcattggga aagaattgtg tgtctgacag
     3721 gttatatgtg aaatacatgt gctggacact gtaacgtgtt gtgttttttg tgaaggtgag
     3781 tttcagtaaa tatccttctc caaaggtgat tgcgagggtg ggctcagaag cattcttttc
     3841 accacaagag ctgctattct tcaggacttc tgcagaggct ggcagggtca tattcgagac
     3901 cttagaaaca tgtccagcct cataggtggt cagaaaggag gcagagaagc tggccattat
     3961 acacgctgtg ccgttgttgt ctttcacctc gaacagtgct ggggcgctgt gtgcaaggcc
     4021 tgccagcagc aacaggagca gcggccgccg ggcgcccggg gccgccatgg tggcgagaat
     4081 tcgaagcttg agctcgagat ctgagtccgg tagcgctagc ggatctgacg gttcactaaa
     4141 ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     4201 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     4261 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     4321 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     4381 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     4441 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     4501 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     4561 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     4621 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     4681 gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga
     4741 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5341 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5401 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5461 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5521 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     5581 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     5641 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     5701 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     5761 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     5821 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     5881 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     5941 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     6001 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     6061 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     6121 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     6181 agccggccac agtcgat
//
