LOCUS       sequence-266324-        8375 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  human VPS33A ORF was inserted into pMXsIP SPGFP-GS Ci2 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8375)
  AUTHORS   Jiang P, Nishimura T, Sakamaki Y, Itakura E, Hatta T, Natsume T, 
            Mizushima N
  TITLE     The HOPS complex mediates autophagosome-lysosome fusion through 
            interaction with syntaxin 17.
  JOURNAL   Mol Biol Cell. 2014 Apr;25(8):1327-37. doi: 10.1091/mbc.E13-08-0447.
            Epub 2014 Feb 19.
  PUBMED    24554770
REFERENCE   2  (bases 1 to 8375)
  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: 
            "10.1091/mbc.E13-08-0447"; journalName: "Mol Biol Cell"; date: 
            "2014-04"; volume: "25"; issue: "8"; pages: "1327-37"
COMMENT     Sequence Label: pMXsIP GFP-VPS33A
FEATURES             Location/Qualifiers
     source          1..8375
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    11..563
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(155..172)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     382..401
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             575..1177
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1074..1094
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1348..1817
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1947..1964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2118..2706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2198..2217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2870..3661,3662..3730))
                     /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     3493..3512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3731..3835)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3865..4458
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4521..4878
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4937..5353
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5283..5305
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5321..5337
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5363..5737
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5675..5691
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(5769..5771,5772..5774,5775..6485)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5769..5771,5772..5774,5775..6485)
                     /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(5814..5835)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6075..6094)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6422..6443
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cccsccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
       61 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
      121 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
      181 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
      241 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
      301 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
      361 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
      421 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
      481 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
      541 ctttgaaaaa cacgatgata atatggccac aaccatggtt accgagtaca agcccacggt
      601 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
      661 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
      721 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
      781 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
      841 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
      901 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
      961 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     1021 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     1081 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     1141 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtc gacgataaaa taaaagattt
     1201 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     1261 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     1321 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     1381 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     1441 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     1501 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     1561 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     1621 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     1681 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     1741 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     1801 tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     1861 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgcgg
     2821 ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3841 ttccccgaaa agtgccacct gcataatgaa agaccccacc tgtaggtttg gcaagctagc
     3901 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag aaaagttcag
     3961 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     4021 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     4081 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     4141 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     4201 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     4261 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     4321 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     4381 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     4441 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     4501 ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc cgattgtcta
     4561 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc
     4621 tggcggaccc gtggtggaac tgacgagttc ggaacacccg gccgcaaccc tgggagacgt
     4681 cccagggact tcgggggccg tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt
     4741 tggactcttt ggtgcacccc cctaatagga gggatatgtg gttctggtag gagacgagaa
     4801 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc
     4861 cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta
     4921 tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct taggtcactg
     4981 gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt
     5041 taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt
     5101 taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca
     5161 cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg
     5221 ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct
     5281 cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc
     5341 ttctctaggc gcccccatat ggccatatga gatcttatat ggggcacccc cgccccttgt
     5401 aaacttccct gaccctgaca tgacaagagt tactaacagc ccctctctcc aagctcactt
     5461 acaggctctc tacttagtcc agcacgaagt ctggagacct ctggcggcag cctaccaaga
     5521 acaactggac cgaccggtgg tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg
     5581 ccgacaccag actaagaacc tagaacctcg ctggaaagga ccttacacag tcctgctgac
     5641 cacccccacc gccctcaaag tagacggcat cgcagcttgg atacacgccg cccacgtgaa
     5701 ggctgccgac cccgggggtg gaccatcctc tagactgccg gatctagcta gttaattaag
     5761 gatctgccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     5821 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     5881 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     5941 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     6001 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     6061 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     6121 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     6181 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     6241 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     6301 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     6361 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     6421 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     6481 acaagggtgg cggtggctcg ggcggtggtg ggtcgggtgg cggcggatct atcgatgaat
     6541 tgaattcacg cgttcgcgag ttaacggatc cctcgagcat ggcggctcat ctgtcctacg
     6601 gccgagtgaa cctaaacgtg ttgcgcgagg cggtgcgtcg cgagctgcgc gagttcctgg
     6661 acaagtgcgc aggaagcaag gcaatagttt gggatgaata cctaactgga ccctttggcc
     6721 tgattgcaca gtattcacta ttgaaggaac atgaagtgga aaaaatgttc acacttaaag
     6781 gaaatcgttt gccggcagct gatgtgaaga atataatttt ttttgtcaga cccaggctag
     6841 agttgatgga tataatcgct gaaaacgtgc tcagtgaaga tagacgaggc ccaacgagag
     6901 attttcatat tctgtttgtg ccacgccgta gcctgttgtg cgaacagcgg ttgaaggatc
     6961 tgggtgtctt gggatccttt attcacaggg aggagtacag cttagatctc attccattcg
     7021 atggggatct cttatccatg gaatcagagg gtgcattcaa agagtgctac ctggagggtg
     7081 accagacgag cctgtaccac gcagccaagg ggctgatgac cctgcaagct ctgtatggaa
     7141 cgatccccca gatctttggg aaaggagaat gcgctcggca agtggccaat atgatgatca
     7201 ggatgaagag agagtttaca ggaagccaga attcaatatt tcctgttttt gataatctct
     7261 tgttgcttga tcggaatgtg gatttattaa cacctcttgc cactcagctg acatatgaag
     7321 gactcattga tgaaatttat ggcattcaga acagttatgt gaaattacct ccagagaaat
     7381 ttgcacctaa gaaacagggc gatggtggta aggacctccc cacggaagca aagaagctgc
     7441 agctgaattc tgcagaggag ctctatgctg agatccgaga taagaacttc aacgcagttg
     7501 gctctgtgct cagcaagaaa gcaaagatca tctctgcagc attcgaggaa agacacaatg
     7561 ctaagaccgt gggggagatc aagcagtttg tttcccagtt gccccacatg caggcagcaa
     7621 ggggctcgct tgcaaaccat acctcaattg cagaattgat caaagatgtc actacttctg
     7681 aagacttttt tgataaatta accgtggaac aggagtttat gtctggaata gacactgata
     7741 aggtcaacaa ttacattgag gattgtatcg cccaaaagca ctcgttgatc aaggtgttaa
     7801 gactagtttg cctccaatcc gtgtgtaata gtgggctcaa acaaaaagtt ttggattatt
     7861 acaaaagaga gattctccag acatacggct atgagcacat attgacctta cacaacctgg
     7921 agaaggccgg cctgctgaaa ccgcagacgg ggggcagaaa caattaccca actatacgga
     7981 aaacattacg cctctggatg gatgatgtta atgagcaaaa ccccacggac atatcgtatg
     8041 tgtacagtgg gtatgccccg ctcagtgtgc ggctggccca gctgctttcc cggcctggct
     8101 ggcggagcat cgaggaggtc ctccgcatcc tcccagggcc ccactttgag gagcggcagc
     8161 cactgcccac aggactgcag aagaaacgtc aaccgggaga aaaccgagtg actctgatat
     8221 ttttccttgg gggcgtaacc ttcgctgaaa ttgctgccct gcgatttctc tcccagttgg
     8281 aagatggagg tacagaatat gtcattgcca ccactaaact aatgaatgga accagttgga
     8341 tagaggctct gatggaaaaa cctttctagg cggcc
//
