LOCUS       sequence-270274-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retrovirus construct for stable expressing mEGFP tagged protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Lazarou M, Sliter DA, Kane LA, Sarraf SA, Wang C, Burman JL, Sideris
            DP, Fogel AI, Youle RJ
  TITLE     The ubiquitin kinase PINK1 recruits autophagy receptors to induce 
            mitophagy.
  JOURNAL   Nature. 2015 Aug 20;524(7565):309-14. doi: 10.1038/nature14893. Epub
            2015 Aug 12.
  PUBMED    26266977
REFERENCE   2  (bases 1 to 6536)
  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"; doi: 
            "10.1038/nature14893"; journalName: "Nature"; date: "2015-08-20- 
            20"; volume: "524"; issue: "7565"; pages: "309-14"
COMMENT     Sequence Label: pBMN-mEGFP-C1
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..358
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             423..839
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     769..791
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     807..823
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    849..1223
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1161..1177
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1232..1234,1235..1237,1238..1948)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1277..1298)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1538..1557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1885..1906
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1949..2014
                     /label=MCS
                     /note="multiple cloning site"
     LTR             2202..2671
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2801..2818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2972..3560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3052..3071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3724..4515,4516..4584))
                     /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     4347..4366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4585..4689)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  4834..5601
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     LTR             6003..6472
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
       61 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
      121 tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg
      181 tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc
      241 ccttagagga gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
      301 cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca
      361 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aatatgggcc
      421 cgggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
      481 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
      541 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
      601 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
      661 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
      721 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
      781 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc
      841 ccatatggcc atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc
      901 ctgacatgac aagagttact aacagcccct ctctccaagc tcacttacag gctctctact
      961 tagtccagca cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac
     1021 cggtggtacc tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta
     1081 agaacctaga acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc
     1141 tcaaagtaga cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg
     1201 ggggtggacc atcctctaga ctgccgccac catggtgagc aagggcgagg agctgttcac
     1261 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1321 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1381 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1441 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1501 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1561 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1621 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1681 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1741 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1801 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     1861 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1921 cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa
     1981 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcacgat
     2041 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     2101 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     2161 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     2221 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     2281 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     2341 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     2401 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     2461 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     2521 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     2581 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     2641 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     2701 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     2761 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3661 agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3721 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3781 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3841 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3901 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3961 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4021 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4081 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4141 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4201 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4261 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4321 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4381 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4441 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4501 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4561 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4621 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4681 ggttccgcgc acatttcccc gaaaagtgcc accagctttg ctcttaggag tttcctaata
     4741 catcccaaac tcaaatatat aaagcatttg acttgttcta tgccctaggg ggcgggggga
     4801 agctaagcca gcttttttta acatttaaaa tgtggtaatg actccaactt attgatagtg
     4861 ttttatgttc agataatgcc cgatgacttt gtcatgcagc tccaccgatt ttgagaacga
     4921 cagcgacttc cgtcccagcc gtgccaggtg ctgcctcaga ttcaggttat gccgctcaat
     4981 tcgctgcgta tatcgcttgc tgattacgtg cagctttccc ttcaggcggg attcatacag
     5041 cggccagcca tccgtcatcc atatcaccac gtcaaagggt gacagcaggc tcataagacg
     5101 ccccagcgtc gccatagtgc gttcaccgaa tacgtgcgca acaaccgtct tccggagact
     5161 gtcatacgcg taaaacagcc agcgctggcg cgatttagcc ccgacatagc cccactgttc
     5221 gtccatttcc gcgcagacga tgacgtcact gcccggctgt atgcgcgagg ttaccgactg
     5281 cggcctgagt tttttaagtg acgtaaaatc gtgttgaggc caacgcccat aatgcgggct
     5341 gttgcccggc atccaacgcc attcatggcc atatcaatga ttttctggtg cgtaccgggt
     5401 tgagaagcgg tgtaagtgaa ctgcagttgc catgttttac ggcagtgaga gcagagatag
     5461 cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag
     5521 ggacagctga tagaaacaga agccactgga gcacctcaaa aacaccatca tacactaaat
     5581 cagtaagttg gcagcatcac cttaaaatgt taattccatt ttaaatgcac agatgttttt
     5641 atttcataag ggtttcaatg tgcatgaatg ctgcaatatt cctgttacca aagctagtat
     5701 aaataaaaat agataaacgt ggaaattact tagagtttct gtcattaacg tttccttcct
     5761 cagttgacaa cataaatgcg ctgctgagaa gccagtttgc atctgtcagg atcaatttcc
     5821 cattatgcca gtcatattaa ttactagtca attagttgat ttttattttt gacatataca
     5881 tgtgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     5941 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
     6001 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     6061 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     6121 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     6181 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     6241 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     6301 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
     6361 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     6421 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     6481 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggt
//
