LOCUS       sequence-429743-        7423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7423)
  AUTHORS   Itakura E, Mizushima N
  TITLE     p62 Targeting to the autophagosome formation site requires 
            self-oligomerization but not LC3 binding.
  JOURNAL   J Cell Biol. 2011 Jan 10;192(1):17-27.
  PUBMED    21220506
REFERENCE   2  (bases 1 to 7423)
  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: "2011-01-10"; volume: "192(1)"; pages: "17-27"
COMMENT     Sequence Label: pMXs-puro GFP-p62 dC
FEATURES             Location/Qualifiers
     source          1..7423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             join(1905..1907,1908..1910,1911..2621)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1905..1907,1908..1910,1911..2621)
                     /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(1950..1971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2211..2230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2558..2579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3547..4121,4122..4124,4125..4133)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3714..3731)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3941..3960
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4134..4736
                     /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     4633..4653
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4907..5376
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5506..5523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5677..6265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5757..5776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6429..7220,7221..7289))
                     /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     7052..7071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagaaaa 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 tgccatggtg agcaagggcg
     1921 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1981 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2041 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2101 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2161 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2221 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2281 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2341 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2401 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2461 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2521 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2581 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggtggcggt ggctcgggcg
     2641 gtggtgggtc gggtggcggc ggatctatcg atgaattgaa ttccatggcg tcgttcacgg
     2701 tgaaggccta tcttctgggc aaggaggagg cgacccgcga gatccgccgc ttcagcttct
     2761 gcttcagccc ggagccggag gcggaagccc aagccgcggc cggcccgggg ccctgcgaga
     2821 ggctgctgag ccgagtggct gtgctgttcc ccacgctgag gcctggcggc ttccaggcgc
     2881 actaccgcga tgaggatggg gacttggttg ccttttccag tgatgaggag ctgacaatgg
     2941 ctatgtccta tgtgaaagat gacatcttcc gcatctacat taaagagaag aaggagtgcc
     3001 ggcgggaaca tcgcccacca tgtgctcagg aggcaccccg aaacatggtg caccccaatg
     3061 tgatctgtga tggttgcaac gggcctgtgg tgggaactcg ctataagtgc agtgtgtgcc
     3121 cagactacga cctgtgcagc gtgtgcgagg ggaagggcct gcacagggaa cacagcaagc
     3181 tcatctttcc caaccccttt ggccacctct ctgatagctt ctctcatagc cgctggcttc
     3241 ggaagctgaa acatggacac tttggctggc ctggctggga gatgggccca ccggggaact
     3301 ggagcccacg tcctcctcgt gcaggggatg gccgcccttg ccctacagct gagtcagctt
     3361 ctgctccacc agaagatccc aatgtcaatt tcctgaagaa tgtgggggag agtgtggcag
     3421 ctgccctcag ccctctaggc attgaggttg acattgatgt ggaacatgga gggaagagat
     3481 gagaatcacg cgttcgcgag ttaacggatc cctcgagcgg ccgctacgta aattccgccc
     3541 cccccccccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3601 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3661 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3721 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3781 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3841 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3901 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3961 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     4021 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     4081 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtta
     4141 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4201 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4261 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4321 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4381 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4441 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4501 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4561 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     4621 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4681 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg
     4741 acgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     4801 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     4861 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     4921 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     4981 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     5041 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     5101 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     5161 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     5221 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     5281 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     5341 ctgagtgatt gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat
     5401 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     5461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6361 cacctagatc cttttgcggc cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg cat
//
