LOCUS       sequence-337178-        8362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To monitor the turnover of SNX9 in lysosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8362)
  AUTHORS   Towers CG, Wodetzki DK, Thorburn J, Smith KR, Caino MC, Thorburn A
  TITLE     Mitochondrial-derived vesicles compensate for loss of LC3-mediated 
            mitophagy.
  JOURNAL   Dev Cell. 2021 Jul 26;56(14):2029-2042.e5. doi: 
            10.1016/j.devcel.2021.06.003. Epub 2021 Jun 24.
  PUBMED    34171288
REFERENCE   2  (bases 1 to 8362)
  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.devcel.2021.06.003"; journalName: "Dev Cell"; date: 
            "2021-07-26- 26"; volume: "56"; issue: "14"; pages: "2029-2042.e5"
COMMENT     Sequence Label: mCherry-GFP-SNX9
FEATURES             Location/Qualifiers
     source          1..8362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..171
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     411..432
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      474..483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(492..516)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(544..1251)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(819..838)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1088..1106
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1248..1257
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1268..1684)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1284..1300)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1316..1338)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    1885..2084
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             2149..2618
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        2649..2753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2754..2822,2823..3614)
                     /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     complement(2972..2991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3774..4362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4263..4282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4516..4533
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             4663..5132
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(5353..5952)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5436..5456)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5933..5952
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5962..6291)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5976..6111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6276..6296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(8123..8362,1..471,472..474,475..477))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(8123..8362,1..471,472..474,475..477))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8165..8186)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
       61 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
      121 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
      181 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
      241 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
      301 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
      361 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
      421 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
      481 ggcgaccgga aagcctgctt ttttgtacaa acttgttcct gggccagcgg atctgagtcc
      541 ggacttgtac agctcgtcca tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg
      601 ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta
      661 ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta
      721 gtggccgccg tccttcagct tcagcctctg cttgatctcg cccttcaggg cgccgtcctc
      781 ggggtacatc cgctcggagg aggcctccca gcccatggtc ttcttctgca ttacggggcc
      841 gtcggagggg aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag
      901 ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca
      961 cttgaagccc tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac
     1021 gtaggccttg gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg
     1081 gccacccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc
     1141 ctcgccctcg atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg
     1201 catgaactcc ttgatgatgg ccatgttatc ctcctcgccc ttgctcacca tggtggcaga
     1261 tccggccggc gcctagagaa ggagtgaggg ctggataaag ggaggatcga ggcggggtcg
     1321 aacgaggagg ttcaaggggg agagacgggg cggatggagg aagaggaggc ggaggcttag
     1381 ggtgtacaaa gggcttgacc cagggagggg ggtcaaaagc caaggcttcc caggtcacga
     1441 tgtaggggac ctggtctggg tgtccatgcg ggccaggtga aaagaccttg atcttaacct
     1501 gggtgatgag gtctcggtta aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc
     1561 attctgcaga gcagaaggta acccaacgtc tcttcttgac atctaccgac tggttgtgag
     1621 cgatccgctc gacatctttc cagtgaccta aggtcaaact taagggagtg gtaacagtct
     1681 ggccctaatt ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc
     1741 tgcagcgctg cagcagacaa gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag
     1801 acggaggcgg gaactgtttt aggttctcgt ctcctaccag aaccacatat cctgacgggg
     1861 tcggattcca catcgactcc cttcctcagg tcgggccaca aaaacggccc ccaaagtccc
     1921 tgggacgtct cccagggttg cggccgggtg ttcagaactc gtcagttcca ccacgggtcc
     1981 gccagataca gagctagtta gctaactagt accgacgcag gcgcataaaa tcagtcatag
     2041 acactagaca atcggacaga cacagataag ttgctggcca gcttgcctcc cggtggtggg
     2101 tcggtggtcc ctgggcaggg gtctcccgat cccggacgag cccccaaatg aaagaccccc
     2161 gctgacgggt agtcaatcac tcagaggaga ccctcccaag gaacagcgag accacaagtc
     2221 ggatgcaact gcaagagggt ttattggata cacgggtacc cgggcgactc agtcaatcgg
     2281 aggactggcg ccccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc
     2341 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt
     2401 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggctgga
     2461 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
     2521 tatcctgttt ggcccatatt cagctgttcc atctgttctt ggccctgagc cggggcagga
     2581 actgcttacc acagatatcc tgtttggccc atattcaggc tgcaggtggc acttttcggg
     2641 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2701 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2761 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2821 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2881 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2941 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3001 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3061 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3121 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3181 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3241 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3301 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3361 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3421 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3481 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3541 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3601 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttgcggc
     3661 cgcaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3721 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3781 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3841 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3901 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3961 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4021 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4081 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4141 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4201 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4261 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4321 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4381 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4441 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4501 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4561 ccgcctctcc ccgcgcgttg gccgattcat taatgcaact atggccattt aatgtaaata
     4621 cttaagaaaa aaaaccaaat taattttgat acatgctgca tgtgaaagac ccccgctgac
     4681 gggtagtcaa tcactcagag gagaccctcc caaggaacag cgagaccaca agtcggatgc
     4741 aactgcaaga gggtttattg gatacacggg tacccgggcg actcagtcaa tcggaggact
     4801 ggcgccccga gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga
     4861 acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc
     4921 cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc tggaccgcat
     4981 ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct
     5041 gtttggccca tattcagctg ttccatctgt tcttggccct gagccggggc aggaactgct
     5101 taccacagat atcctgtttg gcccatattc agctgttcca tctgttcctg accttgatct
     5161 gaacttctct attctcagtt atgtattttt ccatgccttg caaaatggcg ttacttaagc
     5221 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
     5281 aatcttttat tttatcgatg catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc
     5341 gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
     5401 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg
     5461 cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
     5521 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag
     5581 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
     5641 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
     5701 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
     5761 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
     5821 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
     5881 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
     5941 gtactcggtc atggtaagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     6001 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     6061 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     6121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     6181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     6241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     6301 ggtcgaccta catcactgga aagcggctga gggcctgcct cagcttttct gcaatcgttt
     6361 cgtaaaattg cacttgctgc tccaggtaca ggcggatgac actgttgtaa tcatagatcc
     6421 ggttactgtg aaagtgattc atctcagctt gcaacgcgta agacatgatg ctgactctct
     6481 tcaccatgtt ctgtttgtct tgtagggtga ttttacttgt tgcaactagt ttgtcacttt
     6541 ctttcacttt ttctattgct cccttgtgag tgccaatgat gtcagggaag cagccaagaa
     6601 aacctttata ctcgtgatta cattccatca ggaaatggag atctttcttt ggctgttctg
     6661 ccacgagact ggcaatttct tcataagtct ttcctgcttc tgttattgca tcattgagat
     6721 ctgtttcacc ttgatagcca ctggaactga acactgtggc caaactctgc aaggcctttc
     6781 ctatcttctg atattccttg ggtaatgggc ccgtgcagcg cttccagtgc tcctgcccca
     6841 ccgtcagcag ctccttcacg ccgtcatcca tggccttggt gaacttcccc acagcctcgc
     6901 acttctgctc tatttctact aagtccaagt caggtgcctc tggttccatg gtggaaaata
     6961 tcatgactcc cgccagctca tctctctcgg ccttcctctt tccagttttc cattccttct
     7021 catctcggaa atttaggaac tgctggaaaa cttcactttc tgagattact ggatggcgac
     7081 acatcctggt catccaggcc tgaagtctct ccatgcgcat tttgataaat tcctcttcaa
     7141 agcggcctgt gacttgtttg tctggaagag aagggattgg aatggctgac ccaaacttaa
     7201 ccaggagacg ctcatataac cagtcaaagt gcttatacct gtggtttaca gatcgattag
     7261 tgttagtagg tgttagctga tattcgatgt agctctttag accatacatt ttggagcctt
     7321 tcctgggatc tgctaccaca cagtcaaaag tagaggtagg ataaacccac attgggccat
     7381 aatctccaac aatgatggga attttctctt tgggttttgc tagttgtttg gccaacaaat
     7441 actgttccgt gccaggtttc gcaaatccag gaaatttgtt aaggggaatt ttcatggatg
     7501 aggaactagc acgactgttt cctcgctgag cgccgcctgc atcagctgac tctgaatcct
     7561 taaagtagga agaggatttg ggcccatccc agtcttcatc ccagtcatca tcatcaccag
     7621 ttgctggtcc ttggtaggcc tgggggtggc cgaaggcagt gtcccagttg ttgggagtgt
     7681 ttgtgtttct ttgggctcca gccccctctg gctgggcccc ccagccttct gagctttccc
     7741 agttcccaga tttggaggca ctccaggctg accaggggtc attgccactg ccaacctggt
     7801 gattgttgct ggcagccgac gaactggcgt gagctgtgct ggctgagaga gaatcaagga
     7861 aggcttggtc agccactgaa tttccacaag aaaattgatc ttttccatca ctgggtaaaa
     7921 tttcaacgta gtctgtggga accagccctc gttctccttt gatgtttctt ccttccagcc
     7981 atcctccacc tacatccgga tttgtgattg tgatgatttc tccttcatta accgtcagtt
     8041 cattatttcc aggttcagca gcaaaatcat acataacccg agccttggtg gccatgaatt
     8101 cggtaccgga tctgagtccg gacttgtaca gctcgtccat gccgagagtg atcccggcgg
     8161 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
     8221 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     8281 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     8341 tgaagttcac cttgatgccg tt
//
