LOCUS       sequence-435465-        7454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Lamp1 fused to mNeon  Green.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  AUTHORS   Tudorica DA, Basak B, Puerta Cordova AS, Khuu G, Rose K, Lazarou M, 
            Holzbaur ELF, Hurley JH
  TITLE     A RAB7A phosphoswitch coordinates Rubicon Homology protein 
            regulation of Parkin-dependent mitophagy.
  JOURNAL   J Cell Biol. 2024 Jul 1;223(7):e202309015. doi: 
            10.1083/jcb.202309015. Epub 2024 May 10.
  PUBMED    38728007
REFERENCE   2  (bases 1 to 7454)
  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: "2024-07-1"; volume: "223(7):e202309015. doi"; pages: 
            " 10.1083/jcb.202309015. Epub 2024 May 10"
COMMENT     Sequence Label: pCIG2-IRES-Lamp1-mNeon Green
FEATURES             Location/Qualifiers
     source          1..7454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(801..1592,1593..1661))
                     /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     1424..1443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1797..2176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2178..2454
                     /label=chicken beta-actin promoter
     intron          2455..3471
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3395..3417
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3479..3498
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    3526..3578
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3580..4153
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3603..4155
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3747..3764)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3974..3993
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4155..4217,4218..5375)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      5430..5439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5436..6146
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      5691..5700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6238..6257)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6303..6358
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6357..6376)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6749..6765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6749..6765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6762..6784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6773..6789
                     /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(6797..6803,6804..6821,6822..6827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6842..6863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6922..7117
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6968..7103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7123..7257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7160..7179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7214..7233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7325..7342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgggggg
     2281 ggcggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
     2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     2461 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     2521 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     2581 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
     2641 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
     2701 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
     2761 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     2821 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     2881 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     2941 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     3001 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     3061 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     3121 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     3181 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     3241 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     3301 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     3361 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3421 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3481 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattgctcga gctcaagctt
     3541 cgaattctgc agtcgacggt accgcgggcc cgggatccgc ccctctccct cccccccccc
     3601 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     3661 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     3721 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     3781 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     3841 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     3901 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     3961 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     4021 ggtaccccat tgtatgggat ctgatctggg gcctcggtac acatgcttta catgtgttta
     4081 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4141 aacacgatga taatatggcg gccccgggcg cccggcgtcc gctgctcctg ttgctgctgg
     4201 caggccttgc acacagcgcc ccagcactgt tcgaggtgaa agacaacaac ggcacagcgt
     4261 gtataatggc cagcttctct gcctcctttc tgaccaccta tgaggctgga catgtttcta
     4321 aggtctcgaa tatgaccctg ccagcctctg cagaagtcct gaagaatagc agctcttgtg
     4381 gtgaaaagaa tgcttctgag cccaccctcg caatcacctt tggagaagga tatttactga
     4441 aactcacctt cacaaaaaac acaacacgtt acagtgtcca gcacatgtat ttcacatata
     4501 acctgtcaga cacacaattc tttcccaatg ccagctccaa agggcccgac actgtggatt
     4561 ccacaactga catcaaggca gacatcaaca aaacataccg atgtgtcagc gacatcaggg
     4621 tctacatgaa gaatgtgacc attgtgctct gggacgctac tatccaggcc tacctgccga
     4681 gtagcaactt cagcaaggaa gagacacgct gcccacagga tcaaccttcc ccaactactg
     4741 ggccacccag cccctcacca ccacttgtgc ccacaaaccc cagtgtgtcc aagtacaatg
     4801 tgactggtga caatggaacc tgcctgctgg cctctatggc actgcaactc aacatcacct
     4861 acatgaagaa ggacaacacg actgtgacca gagcattcaa catcaaccca agtgacaaat
     4921 atagtgggac ttgcggtgcc cagttggtga ccctgaaggt ggggaacaag agcagagtcc
     4981 tggagctgca gtttgggatg aatgccactt ctagcctgtt tttcctgcaa ggagttcagt
     5041 tgaacatgac tcttcctgat gccatagagc ccacgttcag cacctccaac tattccctga
     5101 aagctcttca ggccagtgtc ggcaactcat acaagtgcaa cagtgaggag cacatctttg
     5161 tcagcaaggc gctcgccctc aatgtcttca gcgtgcaagt ccaggctttc agggtagaaa
     5221 gtgacaggtt tgggtctgtg gaagagtgtg tacaggacgg taacaacatg ctgatcccca
     5281 ttgctgtggg cggggccctg gcagggctgg tcctcatcgt cctcatcgcc tacctcatcg
     5341 gcaggaagag gagtcacgcg ggctatcaga ccatctcgga attcggctcc accggctcca
     5401 ccggctccac cggcgcggat ccaccggtcg ccaccatggt gagcaagggc gaggaggata
     5461 acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc aacggtgtgg
     5521 actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag gagttaaacc
     5581 tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct catatcgggt
     5641 atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag gccgccatgg
     5701 tagatggctc cggctaccaa gtccatcgca caatgcagtt tgaagatggt gcctccctta
     5761 ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc caggtgaagg
     5821 ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct gcggactggt
     5881 gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt aagtggagtt
     5941 acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac acctttgcca
     6001 agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt aagacggagc
     6061 tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt accgatgtga
     6121 tgggcatgga cgagctgtac aagtaaagcg gccgcaattc actcctcagg tgcaggctgc
     6181 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     6241 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     6301 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     6361 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     6421 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     6481 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     6541 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     6601 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     6661 tgtccctctt ctcttatgga gatccctcga cctgcaccgt cgaccagctg gtcgacggtg
     6721 caccgtcgac cagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6781 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6841 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6901 acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6961 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7021 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     7081 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct
     7141 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7201 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc
     7261 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7321 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7381 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7441 gagcaaaagg ccag
//
