LOCUS       sequence-402681-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is the Lazarou lab's standard retrovirus vector backbone for 
            expression of mEGFP-tag fusions onto the N-terminus of cloned genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Nguyen TN, Sawa-Makarska J, Khuu G, Lam WK, Adriaenssens E, 
            Fracchiolla D, Shoebridge S, Bernklau D, Padman BS, Skulsuppaisarn 
            M, Lindblom RSJ, Martens S, Lazarou M
  TITLE     Unconventional initiation of PINK1/Parkin mitophagy by Optineurin.
  JOURNAL   Mol Cell. 2023 May 18;83(10):1693-1709.e9. doi: 
            10.1016/j.molcel.2023.04.021.
  PUBMED    37207627
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: 
            "10.1016/j.molcel.2023.04"; journalName: "Mol Cell"; date: 
            "2023-05-18- 18"; volume: "83"; issue: "10"; pages: "1693-1709.e9"
COMMENT     Sequence Label: pBMN mEGFP
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  615..1382
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     promoter        1527..1631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1632..1700,1701..2492)
                     /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     complement(1850..1869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2656..3244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3145..3164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3398..3415
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             3545..4014
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4202..4267
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(join(4268..4978,4979..4981,4982..4984))
                     /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(4310..4331)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4659..4678
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4918..4939
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4981..4990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4993..5367
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(5039..5055)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(5377..5793)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5393..5409)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5425..5447)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5858..6215
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             join(6280..6536,1..213)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 ccttggggca ccctggaaac atctgatggt tctctagaaa ctgctgaggg ctggaccgca
       61 tctggggacc atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc
      121 tgtttggccc atattcagct gttccatctg ttcttggccc tgagccgggg caggaactgc
      181 ttaccacaga tatcctgttt ggcccatatt cagctgttcc atctgttcct gaccttgatc
      241 tgaacttttc tattctcagt tatgtatttt tccatgcctt gcaaaatggc gttacttaag
      301 ctagcttgcc aaacctacag gtggggtctt tcacatgtat atgtcaaaaa taaaaatcaa
      361 ctaattgact agtaattaat atgactggca taatgggaaa ttgatcctga cagatgcaaa
      421 ctggcttctc agcagcgcat ttatgttgtc aactgaggaa ggaaacgtta atgacagaaa
      481 ctctaagtaa tttccacgtt tatctatttt tatttatact agctttggta acaggaatat
      541 tgcagcattc atgcacattg aaacccttat gaaataaaaa catctgtgca tttaaaatgg
      601 aattaacatt ttaaggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga
      661 ggtgctccag tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg
      721 gtgcgtaacg gcaaaagcac cgccggacat cagcgctatc tctgctctca ctgccgtaaa
      781 acatggcaac tgcagttcac ttacaccgct tctcaacccg gtacgcacca gaaaatcatt
      841 gatatggcca tgaatggcgt tggatgccgg gcaacagccc gcattatggg cgttggcctc
      901 aacacgattt tacgtcactt aaaaaactca ggccgcagtc ggtaacctcg cgcatacagc
      961 cgggcagtga cgtcatcgtc tgcgcggaaa tggacgaaca gtggggctat gtcggggcta
     1021 aatcgcgcca gcgctggctg ttttacgcgt atgacagtct ccggaagacg gttgttgcgc
     1081 acgtattcgg tgaacgcact atggcgacgc tggggcgtct tatgagcctg ctgtcaccct
     1141 ttgacgtggt gatatggatg acggatggct ggccgctgta tgaatcccgc ctgaagggaa
     1201 agctgcacgt aatcagcaag cgatatacgc agcgaattga gcggcataac ctgaatctga
     1261 ggcagcacct ggcacggctg ggacggaagt cgctgtcgtt ctcaaaatcg gtggagctgc
     1321 atgacaaagt catcgggcat tatctgaaca taaaacacta tcaataagtt ggagtcatta
     1381 ccacatttta aatgttaaaa aaagctggct tagcttcccc ccgcccccta gggcatagaa
     1441 caagtcaaat gctttatata tttgagtttg ggatgtatta ggaaactcct aagagcaaag
     1501 ctggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2521 tttagattga tttgcggccg gccgcaaaag gatctaggtg aagatccttt ttgataatct
     2581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2761 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3301 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3361 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3421 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcaa
     3481 ctatggccat ttaatgtaaa tacttaagaa aaaaaaccaa attaattttg atacatgctg
     3541 catgtgaaag acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac
     3601 agcgagacca caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg
     3661 cgactcagtc aatcggagga ctggcgcccc gagtgagggg ttgtgggctc ttttattgag
     3721 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata
     3781 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa
     3841 actgctgagg gctggaccgc atctggggac catctgttct tggccctgag ccggggcagg
     3901 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcttggcc
     3961 ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat tcagctgttc
     4021 catctgttcc tgaccttgat ctgaacttct ctattctcag ttatgtattt ttccatgcct
     4081 tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc
     4141 cctttttctg gagactaaat aaaatctttt attttatcgt gatcagttat ctagatccgg
     4201 tggatcccgg gcccgcggta ccgtcgactg cagaattcga agcttgagct cgagatctga
     4261 gtccggactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     4321 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca
     4381 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     4441 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     4501 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     4561 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     4621 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     4681 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     4741 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     4801 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     4861 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     4921 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     4981 ccatggtggc ggcagtctag aggatggtcc acccccgggg tcggcagcct tcacgtgggc
     5041 ggcgtgtatc caagctgcga tgccgtctac tttgagggcg gtgggggtgg tcagcaggac
     5101 tgtgtaaggt cctttccagc gaggttctag gttcttagtc tggtgtcggc ggacccacac
     5161 tgtgtcgccg actcggtaag ggtgaggtac caccggtcgg tccagttgtt cttggtaggc
     5221 tgccgccaga ggtctccaga cttcgtgctg gactaagtag agagcctgta agtgagcttg
     5281 gagagagggg ctgttagtaa ctcttgtcat gtcagggtca gggaagttta caaggggcgg
     5341 gggtgcccca tataagatct catatggcca tatgggggcg cctagagaag gagtgagggc
     5401 tggataaagg gaggatcgag gcggggtcga acgaggaggt tcaaggggga gagacggggc
     5461 ggatggagga agaggaggcg gaggcttagg gtgtacaaag ggcttgaccc agggaggggg
     5521 gtcaaaagcc aaggcttccc aggtcacgat gtaggggacc tggtctgggt gtccatgcgg
     5581 gccaggtgaa aagaccttga tcttaacctg ggtgatgagg tctcggttaa aggtgccgtc
     5641 tcgcggccat ccgacgttaa aggttggcca ttctgcagag cagaaggtaa cccaacgtct
     5701 cttcttgaca tctaccgact ggttgtgagc gatccgctcg acatctttcc agtgacctaa
     5761 ggtcaaactt aagggagtgg taacagtctg gcccgggccc atattttcag acaaatacag
     5821 aaacacagtc agacagagac aacacagaac gatgctgcag cagacaagac gcgcggcgcg
     5881 gcttcggtcc caaaccgaaa gcaaaaattc agacggaggc gggaactgtt ttaggttctc
     5941 gtctcctacc agaaccacat atccctcctc taaggggggt gcaccaaaga gtccaaaacg
     6001 atcgggattt ttggactcag gtcgggccac aaaaacggcc cccgaagtcc ctgggacgtc
     6061 tcccagggtt gcggccgggt gttccgaact cgtcagttcc accacgggtc cgccagatac
     6121 agagctagtt agctaactag taccgacgca ggcgcataaa atcagtcata gacactagac
     6181 aatcggacag acacagataa gttgctggcc agcttacctc ccggtggtgg gtcggtggtc
     6241 cctgggcagg ggtctcccga tcccggacga gcccccaaat gaaagacccc cgctgacggg
     6301 tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt cggatgcaac
     6361 tgcaagaggg tttattggat acacgggtac ccgggcgact cagtcaatcg gaggactggc
     6421 gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag cgcgcgaaca
     6481 gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt tcaggt
//
