LOCUS       sequence-240913-        9527 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 9527)
  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 9527)
  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-IP GFP-NBR1
FEATURES             Location/Qualifiers
     source          1..9527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..375
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     313..329
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(407..409,410..412,413..1123)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(407..409,410..412,413..1123)
                     /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(452..473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(713..732)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1060..1081
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(4153..4727,4728..4730,4731..4739)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4320..4337)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4547..4566
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4740..5342
                     /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     5239..5259
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5513..5982
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6112..6129)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6283..6871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6363..6382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7035..7826,7827..7895))
                     /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     7658..7677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7896..8000)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             8030..8623
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    8686..9043
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             9102..9518
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     9448..9470
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     9486..9502
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga ccctgacatg
       61 acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta cttagtccag
      121 cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg accggtggta
      181 cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac taagaaccta
      241 gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc cctcaaagta
      301 gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga
      361 ccatcctcta gactgccgga tctagctagt taattaagga tctgccatgg tgagcaaggg
      421 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      481 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      541 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      601 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      661 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      721 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      781 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      841 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
      901 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      961 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1021 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1081 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtggcg gtggctcggg
     1141 cggtggtggg tcgggtggcg gcggatctat cgatgaattg aattcacgcg ttcgcgagtt
     1201 aacggatctt atggaaccac aggttactct aaatgtgact tttaaaaatg aaattcaaag
     1261 ctttctggtt tctgatccag aaaatacaac ttgggctgat atcgaagcta tggtaaaagt
     1321 ttcatttgat ctgaatacta ttcaaataaa atacctggat gaggaaaatg aagaggtatc
     1381 catcaacagt caaggagaat atgaagaagc gcttaagatg gcagttaaac agggaaacca
     1441 actgcagatg caagtccacg aagggcacca tgtcgttgat gaagccccac ccccagttgt
     1501 aggagcaaaa cgactagctg ccagggcagg gaagaagcca cttgcacatt actcttcact
     1561 ggtgagagtc ttgggatcag acatgaagac cccagaggat cctgcagtgc agtcgtttcc
     1621 acttgttcca tgtgacacag accagcctca agacaagccc ccagactggt tcacaagcta
     1681 cctggagacg ttcagagaac aagtggttaa cgaaacggtt gagaagcttg aacagaaatt
     1741 acatgaaaag cttgtcctcc agaacccatc cttgggttct tgtccctcag aagtctcaat
     1801 gcctacttca gaagaaacat tgtttttgcc agaaaaccag ttcagctggc atattgcttg
     1861 caacaactgc caaagaagga ttgttggtgt ccgctaccag tgtagcctat gcccatccta
     1921 caatatctgt gaagattgtg aagcagggcc atatggccat gacactaacc acgtcctgct
     1981 gaagttgcgg agacctgttg tgggctcctc tgaaccgttc tgtcactcaa agtactctac
     2041 tcctcgtctt cctgctgctc tggaacaagt caggctccag aaacaggttg ataagaactt
     2101 tcttaaagca gaaaagcaaa ggttgcgagc tgagaagaaa caacgtaaag cagaggtcaa
     2161 ggaacttaaa aagcagctta aactccatag gaaaattcac ctgtggaatt caatccatgg
     2221 actccagagc cccaagtctc ctttaggccg acctgagagc ttgctccagt ctaataccct
     2281 gatgctccct ttgcagccct gtacctccgt tatgccaatg ctcagtgcag catttgtgga
     2341 tgagaatttg cctgatggga ctcaccttca gccaggaacc aagtttatca aacactggag
     2401 gatgaaaaat acaggaaatg taaagtggag tgcagacaca aagctcaagt tcatgtgggg
     2461 aaacctgact ttggcttcca cagaaaagaa ggatgttttg gttccctgcc tcaaggccgg
     2521 ccatgtggga gttgtatctg tggagttcat tgccccagcc ttggagggaa cgtatacttc
     2581 ccattggcgt ctttctcaca aaggccagca atttgggcct cgggtctggt gcagtatcat
     2641 agtagatcct ttcccctccg aagagagccc tgataacatt gaaaagggca tgatcagctc
     2701 aagcaaaact gatgatctca cctgccagca agaggaaact tttcttctgg ctaaagaaga
     2761 aagacagctt ggtgaagtga ctgagcagac agaagggaca gcagcctgca tcccacagaa
     2821 ggcaaaaaat gttgccagtg agagggagct ctacatccca tctgtggatc ttctgactgc
     2881 ccaggacctg ctgtcctttg agctgttgga tataaacatt gttcaagagt tggagagagt
     2941 gccccacaac acccctgtgg atgtgactcc ctgcatgtct cctctgccac atgacagtcc
     3001 tttaatagag aagccaggct tggggcagat agaggaagag aatgaagggg caggatttaa
     3061 agcacttcct gattctatgg tgtcagtaaa gaggaaggct gagaacattg cttctgtgga
     3121 ggaagcagaa gaagacctga gtgggaccca gtttgtgtgt gagacagtaa tccgatccct
     3181 taccttggat gctgccccag accacaaccc tccttgcaga cagaagtcct tgcagatgac
     3241 atttgccttg cctgaaggac cacttggaaa tgagaaggag gagattatcc atatcgctga
     3301 ggaagaagct gtcatggagg aggaggagga tgaggaggat gaggaggagg aggatgagct
     3361 caaagatgaa gttcaaagtc agtcctctgc ttcctcagag gattacatca tcatcctgcc
     3421 tgagtgcttt gataccagcc gccccctggg ggattctatg tacagctctg cgctctcaca
     3481 gccaggcctg gagcgaggtg ctgaaggcaa gcctggggtt gaggctgggc aggaaccagc
     3541 tgaggctggg gaaagactcc ctggagggga gaaccagcca caggagcaca gcataagtga
     3601 catcctcacg acctcacaga ctctggaaac agtgccccta atcccagagg tagtggagct
     3661 tccaccgtca ctgcccagga gctctccttg tgtacatcat catggttccc caggagtgga
     3721 tttaccagtt accataccag aagtttcttc agtccctgat cagatcagag gagagcccag
     3781 aggctcatca ggacttgtaa acagcagaca gaagagctat gaccactcaa ggcaccatca
     3841 tgggagcagc attgctggag gactggtgaa gggggctttg tctgttgctg cctctgcata
     3901 caaggccctg tttgctgggc caccagtcac tgcacagcca ataatttctg aagatcagac
     3961 agcagccctg atggcccatc tctttgaaat gggattctgt gacaggcagc tgaacctacg
     4021 gctgctgaag aaacacaatt acaatatcct gcaggttgtg acagaacttc ttcagttaaa
     4081 caacaacgac tggtacagcc aacgctattg aagatccctc gagcggccgc tacgtaaatt
     4141 ccgccccccc cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     4201 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     4261 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     4321 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     4381 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     4441 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     4501 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     4561 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     4621 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     4681 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     4741 tggttaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     4801 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     4861 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4921 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4981 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5041 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5101 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5161 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5221 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5281 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     5341 gagtcgacga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     5401 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     5461 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     5521 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     5581 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     5641 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     5701 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     5761 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     5821 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     5881 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     5941 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa
     6001 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6961 gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa
     7021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcata atgaaagacc
     8041 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     8101 cataactgag aatagaaaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     8161 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     8221 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     8281 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     8341 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     8401 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     8461 cacaacccct cactcggcgc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     8521 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     8581 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
     8641 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac
     8701 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
     8761 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttcggaac
     8821 acccggccgc aaccctggga gacgtcccag ggacttcggg ggccgttttt gtggcccgac
     8881 ctgagtccaa aaatcccgat cgttttggac tctttggtgc acccccctaa taggagggat
     8941 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     9001 tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt
     9061 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     9121 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     9181 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     9241 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     9301 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     9361 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     9421 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     9481 cctcccttta tccagccctc actccttctc taggcgcccc catatgg
//
