LOCUS       sequence-228102-        7761 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 7761)
  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 7761)
  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
FEATURES             Location/Qualifiers
     source          1..7761
                     /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             1079..1495
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1425..1447
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1463..1479
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1505..1879
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1817..1833
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(1896..1898,1899..1901,1902..2612)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1896..1898,1899..1901,1902..2612)
                     /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(1941..1962)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2202..2221)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2549..2570
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4075..4095)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4080..4409
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4260..4395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4419..5018
                     /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(4419..4438)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4915..4935
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5245..5714
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5844..5861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6015..6603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6095..6114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6767..7558,7559..7627))
                     /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     7390..7409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7628..7732)
                     /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 cacccccctt
      901 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg
     1081 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1141 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1201 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1261 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1321 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1381 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1441 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     1501 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     1561 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     1621 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     1681 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     1741 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     1801 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     1861 tggaccatcc tctagactgc cggatcatcg atgccatggt gagcaagggc gaggagctgt
     1921 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1981 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2041 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2101 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2161 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2221 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2281 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2341 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2401 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2461 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2521 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2581 ggatcactct cggcatggac gagctgtaca agggtggcgg tggctcgggc ggtggtgggt
     2641 cgggtggcgg cggatctatc gatgaattga attccatggc gtcgttcacg gtgaaggcct
     2701 atcttctggg caaggaggag gcgacccgcg agatccgccg cttcagcttc tgcttcagcc
     2761 cggagccgga ggcggaagcc caagccgcgg ccggcccggg gccctgcgag aggctgctga
     2821 gccgagtggc tgtgctgttc cccacgctga ggcctggcgg cttccaggcg cactaccgcg
     2881 atgaggatgg ggacttggtt gccttttcca gtgatgagga gctgacaatg gctatgtcct
     2941 atgtgaaaga tgacatcttc cgcatctaca ttaaagagaa gaaggagtgc cggcgggaac
     3001 atcgcccacc atgtgctcag gaggcacccc gaaacatggt gcaccccaat gtgatctgtg
     3061 atggttgcaa cgggcctgtg gtgggaactc gctataagtg cagtgtgtgc ccagactacg
     3121 acctgtgcag cgtgtgcgag gggaagggcc tgcacaggga acacagcaag ctcatctttc
     3181 ccaacccctt tggccacctc tctgatagct tctctcatag ccgctggctt cggaagctga
     3241 aacatggaca ctttggctgg cctggctggg agatgggccc accggggaac tggagcccac
     3301 gtcctcctcg tgcaggggat ggccgccctt gccctacagc tgagtcagct tctgctccac
     3361 cagaagatcc caatgtcaat ttcctgaaga atgtggggga gagtgtggca gctgccctca
     3421 gccctctagg cattgaggtt gacattgatg tggaacatgg agggaagaga agccgcctga
     3481 cacccactac cccagaaagt tccagcacag gcacagaaga caagagtaac actcagccaa
     3541 gcagctgctc ttcggaagtc agcaaacctg acggggctgg ggagggccct gctcagtctc
     3601 tgacagagca aatgaaaaag atagccttgg agtcggtggg acagccagag gaacagatgg
     3661 agtcgggaaa ctgctcagga ggagacgatg actggacaca tttgtcttca aaagaagtgg
     3721 acccatctac aggtgaactc cagtctctac agatgccaga atcggaaggg ccaagctctc
     3781 tagacccctc acaggaagga cccacagggc tgaaggaagc tgccctatac ccacatctcc
     3841 caccagaggc tgatccccgg ctgattgagt ccctctccca gatgctgtcc atgggtttct
     3901 cggatgaagg cggctggctc accaggctcc tacagaccaa gaattacgac atcggggctg
     3961 ctctggacac gatccagtat tcgaagcacc ctccaccatt gtgagaattc tgcagtcgac
     4021 ggtaccgcgg gcccgggatc cctcgagcgg ccgccagcac agtggtcgac ctgtggaatg
     4081 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4141 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4201 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4261 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4321 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4381 gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac aagcccacgg
     4441 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4501 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4561 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4621 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4681 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4741 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4801 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4861 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4921 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4981 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc
     5041 gcccgaccga aaggagcgca cgaccccatg catcgtcgac gataaaataa aagattttat
     5101 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     5161 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     5221 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     5281 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     5341 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     5401 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     5461 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     5521 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     5581 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     5641 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     5701 gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt
     5761 tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg
     5821 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg
     6721 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6841 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6961 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7201 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7261 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7441 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7621 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7681 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7741 cccgaaaagt gccacctgca t
//
