LOCUS       sequence-256718-        8853 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8853)
  AUTHORS   Hara T, Takamura A, Kishi C, Iemura S, Natsume T, Guan JL, Mizushima
            N
  TITLE     FIP200, a ULK-interacting protein, is required for autophagosome 
            formation in mammalian cells.
  JOURNAL   J Cell Biol. 2008 May 5. 181(3):497-510.
  PUBMED    18443221
REFERENCE   2  (bases 1 to 8853)
  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. 2008 May 5. 181(3):497-510."
COMMENT     Sequence Label: pMXs-IP-EGFP-hFIP200
FEATURES             Location/Qualifiers
     source          1..8853
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     LTR             1..594
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    657..1014
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1073..1489
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1419..1441
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1457..1473
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1499..1873
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1811..1827
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1959..1968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1965..1967,1968..1970,1971..2681)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1965..1967,1968..1970,1971..2681)
                     /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(2010..2031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2271..2290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2618..2639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5399..5410
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5642..5653
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    join(7616..8190,8191..8193,8194..8202)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7783..7800)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8010..8029
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8203..8805
                     /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     8702..8722
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
      121 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      421 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
      901 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1081 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1141 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1201 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1261 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1321 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1381 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1441 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     1501 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     1561 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     1621 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     1681 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     1741 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     1801 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     1861 atcctctaga ctgccggatc tagctagtta attaaggatc ccagtgtggt ggtacgggaa
     1921 ttcctgcagg cctcgagggc cggcgcgccg cggccgccgc caccatggtg agcaagggcg
     1981 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2041 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2101 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2161 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2221 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2281 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2341 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2401 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2461 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2521 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2581 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2641 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag
     2701 ctcaagcttc gaattctatg aagttatatg tatttctggt taacactgga actactctaa
     2761 catttgacac tgaacttaca gtgcaaactg tggcagacct taagcatgcc attcaaagca
     2821 aatacaagat tgctattcaa caccaggtgc tggtggtcaa tggaggagaa tgcatggctg
     2881 cagatcgaag agtgtgtacc tacagtgctg ggacggatac aaatccaatt tttcttttta
     2941 acaaagaaat gatcttatgt gatcgtccac ctgctattcc taaaactacc ttttcgacag
     3001 aaaatgacat ggaaataaaa gttgaagaat ctcttatgat gcctgcagtt tttcatactg
     3061 ttgcttcaag gacacagctt gcattggaaa tgtatgaagt tgccaagaaa ctttgttctt
     3121 tttgtgaagg tcttgtacat gatgaacatc ttcaacacca aggctgggct gcaatcatgg
     3181 ccaacctgga ggactgttca aattcatacc aaaagctact tttcaagttt gaaagtattt
     3241 attcaaatta tctgcagtcc atagaagaca tcaagttaaa acttactcat ttaggaactg
     3301 cagtttcagt aatggccaag attccactgt tggagtgcct aaccagacat agttacagag
     3361 aatgtttggg aagactggat tctttacctg aacatgaaga ctcagaaaaa gctgagacga
     3421 aaagatccac tgaactggtg ctctctcctg atatgcctag aacaactaac gaatctttgt
     3481 taacctcatt tcccaagtca gtggaacatg tgtccccaga taccgcagat gctgaaagtg
     3541 gcaaagaaat tagggaatct tgtcaaagta ctgttcatca gcaagatgaa actacgattg
     3601 acactaaaga tggtgatctg ccctttttta atgtctcttt gttagactgg ataaatgttc
     3661 aagatagacc taatgatgtg gaatctttgg tcaggaagtg ctttgattct atgagcaggc
     3721 ttgatccaag gattattcga ccatttatag cagaatgccg tcaaactatt gccaaacttg
     3781 ataatcagaa tatgaaagcc attaaaggac ttgaagatcg gctctacgcc ctggaccaga
     3841 tgattgctag ctgtggccga ctggtgaatg aacagaaaga gcttgctcag ggatttttag
     3901 ctaatcagaa gagagctgaa aacttaaagg atgcatctgt attacctgat ttatgcctga
     3961 gtcacgcaaa tcagttgatg attatgttgc aaaatcatag aaaactgtta gatattaagc
     4021 agaagtgtac cactgccaaa caagaactag caaataacct acatgtcaga ctgaagtggt
     4081 gttgctttgt aatgcttcat gctgatcaag atggagagaa gttacaagct ttgctccgcc
     4141 tcgtaataga gctgttagaa agagtcaaaa ttgttgaagc tcttagtaca gttcctcaga
     4201 tgtactgctt agctgttgtt gaggttgtaa gaagaaaaat gttcataaaa cactacaggg
     4261 agtgggctgg tgctttagtc aaagatggaa agagattata tgaagcagaa aaatcaaaaa
     4321 gggaatcctt tgggaaatta tttaggaagt cttttttaag aaatcgtctg tttaggggac
     4381 tggactcctg gcccccttcc ttttgtactc aaaagcctcg aaagtttgac tgtgaacttc
     4441 cagatatttc attaaaagat ttacagtttc tgcaatcatt ttgtccttcg gaagttcagc
     4501 cattcctcag ggttccctta ctttgtgact ttgaacctct acaccagcat gtacttgctc
     4561 tacataattt ggtaaaagca gcacaaagtt tggatgaaat gtcacagacc attacagatc
     4621 tactgagtga acaaaaggca tctgtgagtc agacatcccc acagtctgct tcttcaccaa
     4681 ggatggaaag tacagcagga attacaacta ctacctcacc gagaactcct ccaccactga
     4741 ctgttcagga tcccttatgt cctgcagttt gtcccttaga agaattatct ccagatagta
     4801 ttgatgcaca tacgtttgat tttgaaacta ttccccatcc aaacatagaa cagactattc
     4861 accaagtttc tttagacttg gattcattag cagaaagtcc tgaatcagat tttatgtctg
     4921 ctgtgaatga gtttgtaata gaagaaaatt tgtcgtctcc taatcctata agtgatccac
     4981 aaagcccaga aatgatggtg gaatcacttt attcatcagt tatcaatgcg atagacagta
     5041 gacgaatgca ggatacaaat gtatgtggta aggaggattt tggagatcat acttctctga
     5101 atgtccagtt ggaaagatgt agagttgttg cccaagactc tcacttcagt atacaaacca
     5161 ttaaggaaga cctttgccac tttagaacat ttgtacaaaa agaacagtgt gacttctcaa
     5221 attcattaaa atgtacagca gtagaaataa gaaacattat tgaaaaagta aaatgttctc
     5281 tggaaataac actaaaagaa aaacatcaaa aagaactact gtctttaaaa aatgaatatg
     5341 aaggtaaact tgacggacta ataaaggaaa ctgaagagaa tgaaaacaaa attaaaaaat
     5401 tgaagggaga gttagtatgc cttgaggagg ttttacaaaa taaagataat gaatttgctt
     5461 tggttaaaca tgaaaaagaa gctgtaatct gcctgcagaa tgaaaaggat cagaagttgt
     5521 tagagatgga aaatataatg cactctcaaa attgtgaaat taaagaactg aagcagtcac
     5581 gagaaatagt gttagaagac ttaaaaaagc tccatgttga aaatgatgag aagttacagt
     5641 tattgagggc agaacttcag tccttggagc aaagtcatct aaaggaatta gaggacacac
     5701 ttcaggttag gcacatacaa gagtttgaga aggttatgac agaccacaga gtttctttgg
     5761 aggaattaaa aaaggaaaac caacaaataa ttaatcaaat acaagaatct catgctgaaa
     5821 ttatccagga aaaagaaaaa cagttacagg aattaaaact caaggtttct gatttgtcag
     5881 acacgagatg caagttagag gttgaacttg cgttgaagga agcagaaact gatgaaataa
     5941 aaattttgct ggaagaaagc agagcccagc agaaggagac cttgaaatct cttcttgaac
     6001 aagagacaga aaatttgaga acagaaatta gtaaactcaa ccaaaagatt caggataata
     6061 atgaaaatta tcaggtgggc ttagcagagc taagaacttt aatgacaatt gaaaaagatc
     6121 agtgtatttc cgagttaatt agtagacatg aagaagaatc taatatactt aaagctgaat
     6181 taaacaaagt aacatctttg cataaccaag catttgaaat agaaaaaaac ctaaaagaac
     6241 aaataattga actgcagagt aaattggatt cagaattgag tgctcttgaa agacaaaaag
     6301 atgaaaaaat tacccaacaa gaagagaaat acgaagctat tatccagaac cttgagaaag
     6361 acagacaaaa attggtcagc agccaggagc aagacagaga acagttaatt cagaagctta
     6421 attgtgaaaa agatgaagct attcagactg ccctaaaaga atttaaattg gagagagaag
     6481 ttgttgagaa agagttatta gaaaaagtta aacatcttga gaatcaaata gcaaaaagtc
     6541 ctgccattga ctctaccaga ggagattctt caagcttagt tgctgaactt caagaaaagc
     6601 ttcaggaaga aaaagctaag tttctagaac aacttgaaga gcaagaaaaa agaaagaatg
     6661 aagaaatgca aaatgttcga acatctttga ttgcggaaca acagaccaat tttaacactg
     6721 ttttaacaag agagaaaatg agaaaagaaa acataataaa tgatcttagt gataagttga
     6781 aaagtacaat gcagcaacaa gaacgggata aagatttgat agagtcactt tctgaagatc
     6841 gagctcgttt gcttgaggaa aagaaaaagc ttgaagaaga agtcagtaag ttgcgcagta
     6901 gcagttttgt tccttcacca tatgtagcta cagccccaga actttatgga gcttgtgcac
     6961 ctgaactccc aggtgaatca gatagatccg ctgtggaaac agcagatgaa ggaagagtgg
     7021 attcagcaat ggagacaagc atgatgtctg tacaagaaaa tattcatatg ttgtctgaag
     7081 aaaaacagcg gataatgctg ttagaacgaa cattgcaatt gaaagaagaa gaaaataaac
     7141 ggttaaatca aagactgatg tctcagagca tgtcttcagt atcttcaagg cattctgaaa
     7201 agatagctat tagagatttt caggtgggag atttggtact catcatccta gacgaacgcc
     7261 atgacaatta tgtgttattt actgttagtc ctactttata ttttctacat tcagagtctc
     7321 tacctgccct ggatctcaaa ccaggtgagg gtgcttcagg tgcatctaga agaccctggg
     7381 tactcggaaa agtaatggaa aaagaatact gtcaagccaa aaaggcacaa aacagattta
     7441 aagttccttt ggggacaaag ttttacagag tgaaagccgt atcatggaat aagaaagtat
     7501 aagaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc
     7561 ataatcagcc ataccacatt tgtaggcggc cgctacgtaa attccgcccc ccccccccct
     7621 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     7681 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     7741 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     7801 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     7861 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     7921 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     7981 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     8041 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     8101 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     8161 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggttac cgagtacaag
     8221 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     8281 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     8341 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     8401 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     8461 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     8521 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     8581 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     8641 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     8701 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     8761 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga cgataaaata
     8821 aaagatttta tttagtctcc agaaaaaggg ggg
//
