LOCUS       sequence-260856-        6703 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 6703)
  AUTHORS   Yoshii SR, Kishi C, Ishihara N, Mizushima N
  TITLE     Parkin mediates proteasome-dependent protein degradation and rupture
            of the outer mitochondrial membrane.
  JOURNAL   J Biol Chem. 2011 Jun 3;286(22):19630-40. Epub 2011 Mar 18.
  PUBMED    21454557
REFERENCE   2  (bases 1 to 6703)
  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 Biol 
            Chem."; date: "2011-06-3"; volume: "286(22)"; pages: "19630-40. Epub
            2011 Mar 18"
COMMENT     Sequence Label: pMXs-IP GFP-Omp25
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(46..48,49..51,52..762)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(46..48,49..51,52..762)
                     /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(91..112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(352..371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     699..720
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(912..1486,1487..1489,1490..1498)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1079..1096)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1306..1325
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1499..2101
                     /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     1998..2018
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2272..2741
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2871..2888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3042..3630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3122..3141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3794..4585,4586..4654))
                     /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     4417..4436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4655..4759)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4789..5382
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5445..5802
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5861..6277
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6207..6229
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6245..6261
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    6287..6661
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     6599..6615
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
ORIGIN
        1 gggaattcct gcaggcctcg agggccggcg cgccgcggcc gcaccatggt gagcaagggc
       61 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      121 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      181 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      241 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      301 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      361 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      421 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      481 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      541 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      601 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      661 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      721 accgccgccg ggatcactct cggcatggac gagctgtaca agccgcggca tcgaggcgac
      781 ggagagccta gtggagttcc tgtagctgtg gtgctgctgc cagtgtttgc ccttaccctg
      841 gtagcagttt gggccttcgt gagataccga aagcagctct gagcggccgc tacgtaaatt
      901 cnnccccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
      961 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     1021 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     1081 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     1141 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     1201 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     1261 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     1321 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     1381 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     1441 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     1501 ggttaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     1561 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     1621 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     1681 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     1741 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     1801 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     1861 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     1921 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     1981 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     2041 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     2101 agtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     2161 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     2221 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     2281 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     2341 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     2401 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     2461 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     2521 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     2581 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     2641 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     2701 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     2761 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     2821 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2881 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2941 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3001 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3061 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3721 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     3781 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3841 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3901 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3961 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4021 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4081 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4141 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4201 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4261 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4321 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4381 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4441 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4501 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4561 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4621 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4681 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4741 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcataa tgaaagaccc
     4801 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     4861 ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     4921 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4981 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     5041 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     5101 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     5161 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     5221 acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     5281 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     5341 tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat
     5401 cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact
     5461 tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac
     5521 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttcggaaca
     5581 cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg tggcccgacc
     5641 tgagtccaaa aatcccgatc gttttggact ctttggtgca cccccctaat aggagggata
     5701 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
     5761 ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg ttctgtgttg
     5821 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     5881 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     5941 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     6001 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     6061 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     6121 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     6181 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     6241 ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat atgagatctt
     6301 atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa gagttactaa
     6361 cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg aagtctggag
     6421 acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc acccttaccg
     6481 agtcggcgac acagtgtggg tccgccgaca ccagactaag aacctagaac ctcgctggaa
     6541 aggaccttac acagtcctgc tgaccacccc caccgccctc aaagtagacg gcatcgcagc
     6601 ttggatacac gccgcccacg tgaaggctgc cgaccccggg ggtggaccat cctctagact
     6661 gccggatcta gctagttaat taaggatccc agtgtggtgg tac
//
