LOCUS       sequence-140131-        8714 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 8714)
  AUTHORS   Itakura E, Mizushima N
  TITLE     Characterization of autophagosome formation site by a hierarchical 
            analysis of mammalian Atg proteins.
  JOURNAL   Autophagy. 2010 Aug;6(6):764-76.
  PUBMED    20639694
REFERENCE   2  (bases 1 to 8714)
  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: 
            "Autophagy."; date: "2010-08"; volume: "6(6)"; pages: "764-76"
COMMENT     Sequence Label: pMXs-puro GFP-DFCP1
FEATURES             Location/Qualifiers
     source          1..8714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1100..1516
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1446..1468
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1484..1500
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1526..1900
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1838..1854
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(1917..1919,1920..1922,1923..2633)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1917..1919,1920..1922,1923..2633)
                     /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(1962..1983)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2223..2242)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2570..2591
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5049..5069)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5054..5383
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5234..5369
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5393..5992
                     /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(5393..5412)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5889..5909
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6219..6688
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6818..6835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6989..7577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7069..7088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7741..8532,8533..8601))
                     /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     8364..8383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8602..8706)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct tagaggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat atgggcccgg gccagactgt taccactccc ttaagtttga ccttaggtca
     1141 ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg
     1201 ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac
     1261 ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg
     1321 acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc
     1381 ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc
     1441 tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac
     1501 tccttctcta ggcgccccca tatggccata tgagatctta tatggggcac ccccgcccct
     1561 tgtaaacttc cctgaccctg acatgacaag agttactaac agcccctctc tccaagctca
     1621 cttacaggct ctctacttag tccagcacga agtctggaga cctctggcgg cagcctacca
     1681 agaacaactg gaccgaccgg tggtacctca cccttaccga gtcggcgaca cagtgtgggt
     1741 ccgccgacac cagactaaga acctagaacc tcgctggaaa ggaccttaca cagtcctgct
     1801 gaccaccccc accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt
     1861 gaaggctgcc gaccccgggg gtggaccatc ctctagactg ccggatcatc gatgccatgg
     1921 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1981 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2041 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2101 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2161 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2221 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2281 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2341 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2401 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2461 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2521 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2581 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagatcgatg
     2641 aattgaattc gatgagtgcc cagacttccc tagcagagaa gggcctaaat ccggggctca
     2701 tgtgccagga gagttatgca tgcagtggga ctgatgaagc tatctttgaa tgtgatgagt
     2761 gttgtagttt gcagtgtctc cgctgtgagg aggaactcca cagacaggag cgccttcgaa
     2821 accatgagcg gataagactc aaagctggcc atgtccctta ttgtgacccc tgcaagggcc
     2881 ccaatgggca ttcgccaggt gttaggcaaa gggcagccgt gaggtgccag acttgcaaaa
     2941 tcaatctctg cctggaatgc cagaagagga ctcattcagg gggtaacaag agaagacacc
     3001 ccattaccgt gtaccttgtc agtaaggtcc aggagtccct ggagggggaa gagatggatg
     3061 aggagaccaa gaggaagaag atgacagaga gggttgtgag cttcctcctg gtagacgaaa
     3121 atgaagagat ccaggtaaca aatgaagagg actttatcag gaaattggac tgtaagcctg
     3181 accagcatct caaggtggtt tccatttttg gaaatactgg tgatggaaag tctcacacac
     3241 tcaaccacac tttcttctat ggccgagaag tcttcaaaac ctcccctgct caggagtcct
     3301 gcacagtggg agtgtgggca gcctatgacc cagtccataa ggtagcagtg atagacacag
     3361 aaggactctt gggggccacc gtgaatctga gccagaggac acggctgctg ctgaaggtcc
     3421 tggccatctc agacttggtc atctatcgaa ctcatgcaga ccggctgcat aatgacctct
     3481 tcaaattcct tggggatgcc tcggaagctt acctgaagca cttcaccaag gagctcaagg
     3541 ccaccactgc acgatgtggc ctggatgtcc ccttatccac cctgggccct gcagttatta
     3601 ttttccatga gactgtgcac actcagcttc tgggttctga tcacccctcc gaggcgcccg
     3661 agaagctcat ccaggaccgc ttcaggaagc tgggccgctt ccccgaagcc tttagctcca
     3721 ttcactacaa gggaacgagg acatacaatc cgcccacaga cttctctggg cttcggcgtg
     3781 ctctggaaca gctactagag aacaacacca ccaggtctcc ccggcaccca ggggtcatct
     3841 tcaaagccct gaaggcacta agtgaccgct tcagcggtga gatccccgat gaccagatgg
     3901 cgcacagttc attttttcca gacgagtact tcacctgctc ctcgctctgc ctcagctgtg
     3961 gggcgggatg taagaacagc atgaaccacg gcaaggaagg ggtgcctcat gaagccaaga
     4021 gtcgctgtag gtactcacac cagtatgaca accgcgtcta tacctgcaag gcctgctatg
     4081 agagaggcaa ggaagtcagc gtggtgccca aaacatctgc ctccactgac tctccctgga
     4141 tggggctggc aaagtatgcc tggtctgggt atgtgatcga gtgtcctaac tgtggggtgg
     4201 tctatcggag ccgccagtac tggtttggga accaagatcc tgtggatacg gtggtgcgga
     4261 cagagattgt ccacgtgtgg cctggaactg atgcatttct gaaggataac aacaatgccg
     4321 cccagcgtct cttggatgga atgaacttta tggctcagtc agtgtctgag ctcagcctcg
     4381 gacccactaa ggctgtgacg tcttggctaa cggatcagat tgcccctgcc tactggaggc
     4441 ccaactccca gatcctgagc tgcaaccagt gtgcgacatc cttcaaggac aacgacacga
     4501 agcatcactg ccgggcctgt ggggagggct tctgtgacag ctgttcatcc aagacccggc
     4561 cggtgcctga gcggggctgg ggacctgcgc ctgtccgcgt gtgtgacagc tgctacgatg
     4621 ccaggaatgt ccagttagat gtgactgagg cgcaggcaga tgatgaaggt ggaacgctga
     4681 ttgcccggaa ggtgggcgag gccgtgcaga acaccttggg agccgtggtg acagctattg
     4741 acataccact aggcctggta aaagatgccg ccaggccggc ctactgggtg cctgaccatg
     4801 agatcctgca ctgccacaac tgccgcaagg agttcagcgt caagctctcc aaacaccact
     4861 gccgagcttg cggacagggc ttctgcgatg agtgctccca tgactgcagg gctgtcccct
     4921 ctcgtggctg ggaccatcca gtccgagtct gcttcaactg caataaaaag cccggtgacc
     4981 tttaagaatt cacgcgttcg cgagttaacg gatccctcga gcggccgcca gcacagtggt
     5041 cgacctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5101 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5221 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5281 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5341 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     5401 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     5461 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     5521 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5581 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5641 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5701 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5761 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5821 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5881 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5941 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     6001 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcgt cgacgataaa
     6061 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     6121 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     6181 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     6241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     6301 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6361 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     6421 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     6481 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     6541 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     6601 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     6661 ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt
     6721 ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg
     6781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7681 tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7861 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8701 tccgcgcaca tttc
//
